JPWO2004112817A1 - Celery family-derived extract and method for producing the same - Google Patents

Abstract

The present invention relates to an extract obtained by extracting a celery family plant or a processed product thereof with a hydrous alcohol, a method for producing the extract, an insulin amount or an insulin containing the extract as an active ingredient A therapeutic or preventive agent for diseases associated with response modulation and / or diabetic complications, an insulin-like agent characterized by containing the extract as an active ingredient, and the extract Food, beverage or feed, an agent for promoting glucose uptake into cells, characterized by containing the extract as an active ingredient, a differentiation-inducing agent for fat cells, characterized by containing the extract as an active ingredient, An aldose reductase inhibitor comprising the extract as an active ingredient is also provided.

Description

  TECHNICAL FIELD The present invention relates to an extract derived from a celery family plant containing a large amount of physiologically active ingredients, a method for producing the same, and a medicine, food, beverage or feed containing the extract.

Ashitaba is a large perennial herb belonging to the Apiaceae family and is known for its various health promoting effects. For example, as the physiological activity of Ashitaba, antibacterial action, antitumor action, gastric acid secretion inhibitory action, anticancer effect, nerve growth factor production enhancing effect, hepatocyte growth factor production enhancing action are known (for example, international publication) No. 01/76614 pamphlet). However, insulin-like actions such as anti-diabetic action and anti-obesity action have not been known so far.
Insulin is a hormone required for normal carbohydrate, protein, and fat metabolism in mammals. Humans with type I diabetes require external insulin administration for survival because insulin, which is a hormone that supports life, is not sufficiently produced. Humans with type II diabetes need to administer insulin and insulin secretagogues to control the inappropriate blood glucose level to an appropriate level due to factors such as insufficient insulin production and insulin resistance. Become. However, among humans with type II diabetes, insulin and insulin secretagogues are administered to diabetic patients whose cause is insulin resistance caused by hyperinsulinemia, insulin receptor abnormalities, and abnormal downstream signals of insulin receptors. However, there may be no therapeutic effect.
In recent years, in order to solve the side effects of insulin and the above-mentioned problems, substances having physiological functions similar to those of insulin (hereinafter sometimes referred to as insulin-like substances) have been developed. It is a like substance (for example, WO 99/51225 pamphlet), and shikonin derived from Shikon (purine) is an insulin-like substance (for example, Kamei R. et al., Biochem. Biophys. Res. Commune). , 2002, Vol. 292, P642-651). Insulin-like substances such as these improve not only type I diabetics but also type II diabetics, and even type II diabetics caused by insulin resistance, by improving physiological symptoms similar to insulin Is expected to be.

An object of the present invention is an extract derived from a celery family plant containing a large amount of physiologically active ingredients, which is safe from natural products, can be easily ingested, a method for producing the same, and a pharmaceutical, food, or beverage containing the extract Or to provide feed.
Hereinafter, when the present invention is outlined, the first invention of the present invention relates to an extract obtained by extracting a celery family plant or a processed product thereof with a hydrous alcohol.
A second invention of the present invention relates to a method for producing an extract derived from an Aceraceae plant or a processed product thereof, which comprises a step of extracting the Aceraceae plant or a processed product thereof with a hydrous alcohol.
In the first and second inventions of the present invention, Acetaba is preferably exemplified as the Apiaceae plant. Examples of the hydrous alcohol include an aqueous ethanol solution having a concentration of 40 (v / v)% or more and less than 100 (v / v)%.
3rd invention of this invention contains the extract of 1st invention of this invention as an active ingredient, The therapeutic agent of the disease accompanying the modulation | alteration of the amount of insulin or an insulin response, and / or the complication of diabetes Or it relates to a preventive agent.
A fourth invention of the present invention relates to an insulin-like agent characterized by containing the extract of the first invention of the present invention as an active ingredient.
5th invention of this invention is related with the foodstuff, drink, or feed characterized by containing the extract of 1st invention of this invention. In the fifth invention of the present invention, the food, beverage or feed is exemplified by a food, beverage or feed for treating or preventing a disease accompanied by modulation of insulin amount or insulin response and / or a complication of diabetes. .
The sixth invention of the present invention relates to a glucose uptake promoter for cells characterized by containing the extract of the first invention of the present invention as an active ingredient.
The seventh invention of the present invention relates to an agent for inducing differentiation into adipocytes, comprising the extract of the first invention of the present invention as an active ingredient.
The eighth invention of the present invention relates to an aldose reductase inhibitor characterized by containing the extract of the first invention of the present invention as an active ingredient.

本発明の抽出物に含まれる以上の４種のカルコン類化合物（４−ハイドロキシデリシン、キサントアンゲロール、ＴＢ１、ＴＢ２）は後述の参考例１及び２に記載のとおり、天然由来のインスリン様活性成分である。さらに、ＴＢ１およびＴＢ２については、アルドースレダクターゼ阻害作用も有しており、糖尿病の合併症に対してその効果が期待されることが、本発明者らにより見出されている（国際公開第２００４／０３１１６５号パンフレット）。
さらにアシタバから得られる本発明の抽出物中には、カルコン類化合物であるキサントアンゲロールＨ、イソババカルコン、ババクロマノールや、フラバノン類化合物であるムンドゥレアフラバノンＡ、イソババチン、プロストラトールＦが含有されており、これらの化合物は後述の実施例１１、参考例１及び２にも示すとおり、アルドースレダクターゼ阻害作用やインスリン様作用を有している。
すなわち、本発明により得られる抽出物は、健常人への糖尿病の予防効果、糖尿病予備軍と呼ばれる初期の糖尿病の人々に対する改善効果、および糖尿病患者でその合併症の症状が見られる人々に対する糖尿病とその合併症の改善効果を有する、総合的な糖尿病改善又は予防用の医薬品、機能性食品素材として極めて有用である。なお、本発明の抽出物の有するインスリン様作用は、細胞へのグルコース取り込み促進作用、脂肪細胞への分化誘導作用を指標にして、後述の実施例５〜７において示されている。また、本発明の抽出物のアルドースレダクターゼ阻害作用は後述の実施例８、９において示されている。
また、上記のＴＢ１、ＴＢ２はアルドースレダクターゼ阻害作用以外にも、神経細胞保護作用、ＮＯ産生抑制作用、インターロイキン産生抑制作用、骨形成促進作用等を有していることから（例えば、国際公開第２００４／０３１１６５号パンフレット参照）、本発明の抽出物はこれらの生理活性を利用した天然由来の医薬品、健康食品素材としても有用である。
なお、本発明においてインスリン様作用とは、インスリンの有する生理活性のうち、少なくとも１つを示すものであれば特に限定はなく、例えば、細胞における糖、アミノ酸の取り込み促進、グリコーゲン、タンパク質合成及び分解抑制などの代謝調節作用のうち少なくとも１つが例示される。また、インスリン様作用の有無については、後述の実施例５又は６に記載の方法により簡便に測定することができる。本発明の有効成分はインスリン様作用を有することから、治療上または予防上、インスリンの使用が有効であるあらゆる疾患に対し治療効果または予防効果を発揮し得る。
また、本発明のセリ科植物由来の抽出物を公知の方法で分画することによって得られる画分や、分画操作を複数回繰り返すことにより得られる画分も本発明の抽出物に包含される。上記の分画手段としては、抽出、分別沈殿、カラムクロマトグラフィー、薄層クロマトグラフィー等が挙げられる。
本発明において、本発明の抽出物の形状は、特に限定はないが、粉状、固形状、液状のいずれの形状であってもよい。粉状とする場合、特に限定はないが、原料より含水アルコールを溶媒として抽出された本発明の抽出物を濃縮し、さらに賦形剤等を添加し、乾燥、粉砕することにより、粉状の本発明の抽出物を得ることができる。また、当該抽出物を公知の方法で造粒して得た粒状の固形物を、本発明の抽出物として使用することもできる。造粒方法としては、特に限定はないが、転動造粒、攪拌造粒、流動層造粒、気流造粒、押出し造粒、圧縮成型造粒、解砕造粒、噴射造粒又は噴霧造粒等が例示される。また、液状の抽出物としては、含水エタノール抽出物そのもの、その濃縮物や希釈物の他、前記の粉状の抽出物を液体、例えば水やアルコール等に溶解して液状としたものが例示される。
また、本発明は、本発明の抽出物を高濃度又は高純度に含有する食品、飲料又は飼料を提供するが、これらは従来の食品、飲料又は飼料と比べて、本発明の食品、飲料又は飼料中に前記のカルコン類化合物やフラバノン類化合物が高濃度及び／又は高純度に含有されていることを意味する。
なお、本発明において、本発明の抽出物を本発明の有効成分と称し、本発明の有効成分を含有するインスリン量またはインスリン応答の変調を伴う疾患、及び／又は糖尿病の合併症の治療剤又は予防剤を、本発明の治療剤又は予防剤と称することがある。また、該治療剤、予防剤の他、インスリン様作用剤を含めて本発明の医薬という場合がある。
本発明に係る有効成分には、後述するように特に毒性は認められない。また、副作用の発生の心配もない。それゆえ、安全かつ適切に疾患の治療又は予防を行うことができる。従って、当該有効成分を含んでなる本発明の治療剤、予防剤、食品、飲料または飼料は、インスリン量またはインスリン応答の変調を伴う疾患、及び／又は糖尿病の合併症の治療または予防に有効である。
本発明において、インスリン量またはインスリン応答の変調を伴う疾患としては、血中のインスリンレベルの変化、インスリンもしくはインスリン受容体の活性のレベルの変化、インスリン受容体の下流シグナルの異常、及びそれらの組み合わせから選択される因子によって特徴づけられる疾患が挙げられ、例えば、糖尿病、肥満症、高血圧、動脈硬化、コカイン禁断症状、鬱血性心不全、健忘症、心臓血管痙攣、大脳血管痙攣、クロム親和性細胞腫、神経節神経芽腫、ハンチントン病、高脂血症が例示される。糖尿病としては、Ｉ型糖尿病、ＩＩ型糖尿病のいずれもが例示される。また、ＩＩ型糖尿病としては、インスリンやインスリン分泌促進薬を投与しても治療効果の見られないようなインスリン抵抗性が要因の疾患についても包含される。
インスリン量又はインスリン応答の変調を伴う疾患は、その発症段階において、インスリン産生量が不足していたり、あるいはインスリン抵抗性が原因となってインスリンによる作用が不十分となることが多い。本発明の有効成分はインスリン様作用を示すことにより、インスリン量又はインスリン応答の変調を伴う疾患の発症を抑制することができることから、当該疾患の予防効果をも期待でき、ひいては糖尿病の合併症の予防効果をも期待できる。
インスリン抵抗性の状態においてはインスリンによるインスリンレセプターからの信号が阻害されており、インスリンの持つ多彩な機能が発揮されず、種々の代謝異常が生じる。本発明に使用される有効成分は、インスリン抵抗性の症状に対してもインスリン様の効果を発揮することができる。すなわち、本発明の予防剤または治療剤を用いることで、インスリン抵抗性が要因の疾患、例えばインスリンやインスリン分泌促進薬を投与しても治療効果の見られないようなＩＩ型糖尿病に対しても治療もしくは予防効果を発揮することができる。また、本発明の有効成分は、血中インスリン量の低下効果をも発揮し得る。すなわち、本発明の医薬を、治療又は予防にインスリン量の低下を要する疾患の治療又は予防剤として使用することもできる。当該疾患としては、特に限定はないが、高インスリン血症やアルツハイマー病等が例示される。また、インスリン受容体を介する刺激と延命効果については密接な関係があるという報告もあることから（Ｓｃｉｅｎｃｅ，ｖｏｌ．２９９，Ｐ５７２〜５７４（２００３年）；Ｎａｔｕｒｅ，ｖｏｌ．４２４，Ｐ２７７〜２８４（２００３年））、本発明の医薬を老化防止剤として使用することもできる。
インスリンは前駆脂肪細胞の脂肪細胞への分化誘導を促進することが知られており、また成熟した脂肪細胞ではグルコースを取り込み、細胞内にトリグリセリドが蓄積されることが知られている（Ｊ．Ｂｉｏｌ．Ｃｈｅｍ．，Ｖｏｌ．２５３，Ｎｏ．２０，Ｐ７５７０〜７５７８（１９７８年））。すなわち、この知見を利用して、インスリンの代わりに被験物質を投与し、脂肪細胞への分化や細胞中のトリグリセリド量を測定することで、被験物質のインスリン様作用を測定することができる。
また、インスリンは細胞へのグルコース取り込み促進作用が知られており、成熟した脂肪細胞ではインスリンの作用により細胞内へのグルコースの取り込みが促進されることが知られている（Ｊ．Ｂｉｏｌ．Ｃｈｅｍ．，Ｖｏｌ．２５３，Ｎｏ．２０，Ｐ７５７９−７５８３（１９７８年））。すなわち、この知見を利用して、インスリンの代わりに被験物質を投与し、成熟脂肪細胞内へのグルコースの取り込み量を測定することで、被験物質のインスリン様作用を測定することができる。
本発明において、糖尿病の合併症としては、例えば、糖尿病性網膜症、糖尿病性末梢神経症、白内障、難聴、知覚異常、筋力の委縮等の糖尿病精神症、腎不全等の糖尿病性腎症、動脈硬化や脳梗塞等の糖尿病性血管疾患、白血球の食菌作用の低下に起因する感染病、糖尿病性昏睡等が例示される。
本発明の治療剤または予防剤としては、本発明に係る前記有効成分を公知の医薬用担体と組み合わせて製剤化したものが挙げられる。また、本発明の治療剤または予防剤としては、前記有効成分を当該有効成分と同じ用途に使用可能な他の成分、例えば、公知のインスリン製剤、インスリン分泌促進剤、インスリン抵抗性改善剤、食後過血糖改善剤、インスリン様作用剤、アルドースレダクターゼ阻害剤などと配合することもできる。
本発明の治療剤または予防剤の製造は、通常、前記有効成分を薬学的に許容できる液状または固体状の担体と配合することにより行われ、所望により溶剤、分散剤、乳化剤、緩衝剤、安定剤、賦形剤、結合剤、崩壊剤、滑沢剤等を加えて、錠剤、顆粒剤、散剤、粉末剤、カプセル剤等の固形剤、通常液剤、懸濁剤、乳剤等の液剤とすることができる。また、使用前に適当な担体の添加によって液状となし得る乾燥品や、その他、外用剤とすることもできる。
医薬用担体は、治療剤または予防剤の投与形態および剤型に応じて選択することができる。固体組成物からなる経口剤とする場合は、錠剤、丸剤、カプセル剤、散剤、細粒剤、顆粒剤等とすることができ、たとえば、デンプン、乳糖、白糖、マンニット、カルボキシメチルセルロース、コーンスターチ、無機塩などが利用される。また経口剤の調製に当っては、更に結合剤、崩壊剤、界面活性剤、潤沢剤、流動性促進剤、矯味剤、着色剤、香料などを配合することもできる。たとえば、錠剤または丸剤とする場合は、所望によりショ糖、ゼラチン、ヒドロキシプロピルセルロースなどの糖衣または胃溶性もしくは腸溶性物質のフィルムで被覆してもよい。液体組成物からなる経口剤とする場合は、薬理学的に許容される乳濁剤、溶液剤、懸濁剤、シロップ剤などとすることができ、たとえば、精製水、エタノールなどが担体として利用される。また、さらに所望により湿潤剤、懸濁剤のような補助剤、甘味剤、風味剤、防腐剤などを添加してもよい。
一方、非経口剤とする場合は、常法に従い本発明の前記有効成分を希釈剤としての注射用蒸留水、生理食塩水、ブドウ糖水溶液、注射用植物油、ゴマ油、落花生油、大豆油、トウモロコシ油、プロピレングリコール、ポリエチレングリコールなどに溶解ないし懸濁させ、必要に応じ、殺菌剤、安定剤、等張化剤、無痛化剤などを加えることにより調製することができる。また、固体組成物を製造し、使用前に無菌水または無菌の注射用溶媒に溶解して使用することもできる。
外用剤としては、経皮投与用または経粘膜（口腔内、鼻腔内）投与用の、固体、半固体状または液状の製剤が含まれる。また、座剤なども含まれる。たとえば、乳剤、ローション剤などの乳濁剤、外用チンキ剤、経粘膜投与用液剤などの液状製剤、油性軟膏、親水性軟膏などの軟膏剤、フィルム剤、テープ剤、パップ剤などの経皮投与用または経粘膜投与用の貼付剤などとすることができる。
以上の各種製剤は、それぞれ公知の医薬用担体などを利用して、適宜、常法により製造することができる。また、かかる製剤における有効成分の含有量は、その投与形態、投与方法などを考慮し、好ましくは後述の投与量範囲で当該有効成分を投与できるような量であれば特に限定されるものではない。本発明の治療剤又は予防剤中の有効成分の含有量としては通常０．１〜１００重量％程度である。
本発明の治療剤又は予防剤は、製剤形態に応じた適当な投与経路で投与される。投与方法も特に限定はなく、内用、外用および注射によることができる。注射剤は、たとえば静脈内、筋肉内、皮下、皮内などに投与し得、外用剤では、たとえば、座剤をその適する投与方法により投与すればよい。
本発明の治療剤または予防剤としての投与量は、その製剤形態、投与方法、使用目的および当該治療剤または予防剤の投与対象である患者の年齢、体重、症状によって適宜設定され一定ではない。一般には、製剤中に含有される前記有効成分の投与量が、乾燥重量で、成人１日当り好ましくは０．００１ｍｇ〜１０ｇ／ｋｇ体重、より好ましくは０．１ｍｇ〜１ｇ／ｋｇ体重となる量である。もちろん投与量は、種々の条件によって変動するので、上記投与量より少ない量で十分な場合もあるし、あるいは範囲を超えて必要な場合もある。投与は、所望の投与量範囲内において、１日内において単回で、または数回に分けて行ってもよい。投与期間も任意である。また、本発明の治療剤または予防剤はそのまま経口投与するほか、任意の飲食品に添加して日常的に摂取させることもできる。
また、本発明は前記有効成分を含むインスリン様作用剤を提供することもできる。当該インスリン様作用剤としては、前記有効成分そのものであってもよく、また、前記有効成分を含む組成物であってもよい。当該インスリン様作用剤は、たとえば、前記有効成分を当該有効成分と同じ用途に使用可能な他の成分、たとえば、公知のインスリン製剤、インスリン分泌促進剤、インスリン抵抗性改善剤、食後過血糖改善剤、インスリン様作用剤などと配合し、上記治療剤または予防剤の製造方法に準じて通常使用される試薬の形態に製造すればよい。当該インスリン様作用剤における前記有効成分の含有量は、当該インスリン様作用剤の投与方法、使用目的などを考慮し、本発明の所望の効果の発現が得られ得るような量であればよく、特に限定されるものではない。典型的なインスリン様作用剤中の有効成分の含有量としては、０．１〜１００重量％程度である。また、当該インスリン様作用剤の使用量も、本発明の所望の効果の発現が得られ得る量であれば特に限定されるものではない。特に、生体に投与して使用する場合には、好ましくは前記治療剤または予防剤における有効成分の投与量の範囲内で有効成分を投与できるような量で使用すればよい。インスリン様作用剤は、インスリン量またはインスリン応答の変調を伴う疾患において有用である。また、当該インスリン様作用剤はインスリン量またはインスリン応答の変調を伴う疾患に対する薬物のスクリーニングにも有用である。さらに当該インスリン様作用剤は、インスリンによる細胞への作用メカニズム研究や、その細胞の物理的変化に関する機能研究にも有用である。また、当該インスリン様作用剤は、血清やインスリン製剤のかわりに、もしくはそれらと共に細胞・組織・臓器培養用の培地に添加して使用することもできる。当該培地は血清やインスリン製剤を低減もしくは含まない細胞・組織・臓器培養用の培地として使用するのに非常に有用である。
また、本発明のインスリン様作用剤をヒトに投与することにより、血中インスリン量の低下を期待することができる。すなわち、本発明のインスリン様作用剤を、治療又は予防にインスリン量の低下を要する疾患の治療又は予防剤として使用することもできる。当該疾患としては、特に限定はないが、高インスリン血症やアルツハイマー病等が例示される。また、インスリン受容体を介する刺激と延命効果については密接な関係があるという報告もあることから（Ｓｃｉｅｎｃｅ，ｖｏｌ．２９９，Ｐ５７２〜５７４（２００３年）；Ｎａｔｕｒｅ，ｖｏｌ．４２４，Ｐ２７７〜２８４（２００３年））、本発明のインスリン様作用剤を老化防止剤として使用することもできる。
本発明に係る有効成分には、後述するように特に毒性は認められない。また、副作用の発生の心配もない。それゆえ、安全かつ適切にインスリン様作用、及び／又はアルドースレダクターゼ阻害作用を生体内で発させることができる。従って、当該有効成分を含んでなる本発明の医薬、食品、飲料または飼料は、インスリン量またはインスリン応答の変調を伴う疾患、及び／又は糖尿病の合併症の治療または予防に有効である。
また、本発明は、本発明の抽出物を含有してなる食品、飲料又は飼料を提供する。本発明の食品、飲料または飼料は、そのインスリン様作用、アルドースレダクターゼ阻害作用により、インスリン量またはインスリン応答の変調を伴う疾患、糖尿病の合併症の症状改善や予防に対して極めて有用である。さらに、本発明の食品又は飲料は、血糖値を低下させる作用を有する、血糖値低下用の食品又は飲料であり、血糖値が気になる方や体脂肪が気になる方に対して有効な機能性食品又は飲料として有用である。
本発明の食品、飲料または飼料は、抗糖尿病作用を有することが知られている他の物質、たとえば、公知のインスリン様作用物質、インスリン分泌促進作用を有する物質、インスリン抵抗性改善作用を有する物質、食後過血糖改善作用を有する物質、アルドースレダクターゼ阻害物質などと配合することもできる。たとえば、難消化性デキストリン等と配合することもできる。
なお、本発明の食品、飲料または飼料において「含有」とは、含有、添加及び／又は希釈を意味する。ここで、「含有」とは食品、飲料または飼料中に本発明の抽出物が含まれるという態様を、「添加」とは食品、飲料または飼料の原料に、本発明の抽出物を添加するという態様を、「希釈」とは本発明の抽出物に、食品、飲料または飼料の原料を添加するという態様をいうものである。
本発明の食品、飲料または飼料の製造法に特に限定はない。たとえば、配合、調理、加工などは一般の食品、飲料または飼料のものに従えばよく、それらの製造法により製造することができ、得られた食品、飲料または飼料に本発明の抽出物が含有されていれば良い。
本発明の食品または飲料としては特に限定はないが、たとえば、本発明の抽出物が含有されてなる、穀物加工品（小麦粉加工品、デンプン類加工品、プレミックス加工品、麺類、マカロニ類、パン類、あん類、そば類、麩、ビーフン、はるさめ、包装餅など）、油脂加工品（可塑性油脂、てんぷら油、サラダ油、マヨネーズ類、ドレッシングなど）、大豆加工品（豆腐類、味噌、納豆など）、食肉加工品（ハム、ベーコン、プレスハム、ソーセージなど）、水産製品（冷凍すりみ、かまぼこ、ちくわ、はんぺん、さつま揚げ、つみれ、すじ、魚肉ハム、ソーセージ、かつお節、魚卵加工品、水産缶詰、つくだ煮など）、乳製品（原料乳、クリーム、ヨーグルト、バター、チーズ、練乳、粉乳、アイスクリームなど）、野菜・果実加工品（ペースト類、ジャム類、漬け物類、果実飲料、野菜飲料、ミックス飲料など）、菓子類（チョコレート、ビスケット類、菓子パン類、ケーキ、餅菓子、米菓類など）、アルコール飲料（日本酒、中国酒、ワイン、ウイスキー、焼酎、ウオッカ、ブランデー、ジン、ラム酒、ビール、清涼アルコール飲料、果実酒、リキュールなど）、嗜好飲料（青汁、緑茶、紅茶、ウーロン茶、コーヒー、清涼飲料、乳酸飲料など）、調味料（しょうゆ、ソース、酢、みりんなど）、缶詰・瓶詰め・袋詰め食品（牛飯、釜飯、赤飯、カレー、その他の各種調理済み食品）、半乾燥または濃縮食品（レバーペースト、その他のスプレッド、そば・うどんの汁、濃縮スープ類）、乾燥食品（即席麺類、即席カレー、インスタントコーヒー、粉末ジュース、粉末スープ、即席味噌汁、調理済み食品、調理済み飲料、調理済みスープなど）、冷凍食品（すき焼き、茶碗蒸し、うなぎかば焼き、ハンバーグステーキ、シュウマイ、餃子、各種スティック、フルーツカクテルなど）、固形食品、液体食品（スープなど）、香辛料類などの農産・林産加工品、畜産加工品、水産加工品などが挙げられる。
本発明の食品は、本発明の抽出物が単独もしくは複数含有、添加および／または希釈されており、特にその形状に限定はなく、タブレット状、顆粒状、カプセル状等の形状の経口的に摂取可能な形状物も包含する。
タブレット状食品の製造方法としては、公知の方法により製造することができ、特に限定はないが、例えばアシタバ乾燥粉末を６０（ｖ／ｖ）％エタノール水溶液で抽出し、次いで濃縮した抽出物に、賦形剤、例えば、デキストリン、難消化性デキストリン、コーンスターチ、タピオカデンプン、サイクロデキストリン等を添加して凍結乾燥、粉砕することにより、アシタバ抽出物の乾燥粉末を得た後、さらに乳糖、結晶セルロース、ショ糖脂肪酸エステル、還元麦芽糖、リン酸カルシウム、卵殻カルシウム、微粒二酸化ケイ素、ＣＭＣ−Ｃａ、トレハロース、ビタミンＣ等の各種ビタミン類、クエン酸、ブドウ糖、砂糖、糖アルコール、ステビア等の甘味料、香料、粉末果汁、海藻由来粉末（例えば、フコイダン含有粉末、アガロオリゴ糖粉末等）、きのこ粉末、乾燥野菜粉末（例えばアシタバ等のセリ科植物粉末）等を適宜混合し、必要に応じて造粒工程を施し、ロータリー式打錠機により打錠して製造することができる。
また、顆粒状食品の製造方法としては、公知の方法により製造することができ、特に限定はないが、例えば上記のタブレット状食品の製造工程においてロータリー式打錠機に施す前に得られる各種混合した粉末に、エタノールを添加して練合し、押出し造粒機により造粒し、これを乾燥させて振動篩で整粒して顆粒状食品を製造することができる。
また、カプセル状食品の製造方法としては、公知の方法により製造することができ、特に限定はないが、例えば上記のタブレット状食品の製造工程においてロータリー式打錠機に施す前に得られる各種混合した粉末を、１号カプセルに充填して製造することができる。また、充填前の粉末にグリセリン脂肪酸エステル、ミツロウ等を添加して乳化させ、ゼラチンとグリセリンを被包材としてソフトカプセルを製造することもできる。
なお、本発明の抽出物はセリ科植物独特の苦味や渋味が低減されており、食品原料としても優れている。すなわち、本発明の食品又は飲料は、嗜好的にも優れた食品又は飲料である。
また、本発明の飲料としては、公知の方法により製造することができ、特に限定はないが、例えばアシタバ乾燥粉末を６０（ｖ／ｖ）％エタノール水溶液で抽出し、濃縮した抽出物に、賦形剤、例えば、デキストリン、難消化性デキストリン、コーンスターチ、タピオカデンプン、サイクロデキストリン等を添加して凍結乾燥し、粉砕することにより、アシタバ抽出物の乾燥粉末を得た後、これに適宜香料、甘味料、果汁、粉末果汁、野菜エキス、野菜ペースト、海藻由来粉末や海藻抽出物（例えば、フコイダン含有物、アガロオリゴ糖含有物等）、きのこ粉末、セリ科植物の乾燥粉末等を添加し、水やアルコールに溶解することにより本発明の飲料を製造することができる。また、アシタバ抽出物の乾燥粉末を水に溶解した後、公知の清涼飲料の製造方法に準じて本発明の飲料を製造することもできる。また、アシタバ抽出物の乾燥粉末の代わりにアシタバ抽出物の濃縮物を使用することもできる。さらに、アシタバの含水アルコール抽出物を用いて、本発明の飲料としてアルコール飲料を製造することもできる。
本発明の食品又は飲料中の前記有効成分の含有量は特に限定されず、その官能と活性発現の観点から適宜選択できるが、例えば、前記有効成分の乾燥重量で、食品中、好ましくは０．００００１重量％以上、より好ましくは０．０００１〜９０重量％、更に好適には０．０００６〜８０重量％であり、例えば、飲料中、好ましくは０．００００１重量％以上、より好ましくは０．０００１〜９０重量％、更に好適には０．０００６〜８０重量％である。また本発明の食品又は飲料は、それらに含有される有効成分が、例えば成人１日当たり、好ましくは０．００１ｍｇ〜１０ｇ／ｋｇ体重、より好ましくは０．１ｍｇ〜１ｇ／ｋｇ体重となるように摂取すればよい。
また、本発明は、本発明の抽出物を含有、添加および／または希釈してなる、生物用の飼料を提供するものであり、さらに、別の一態様として、本発明の抽出物を生物に投与することを特徴とする生物の飼育方法をも提供する。また、本発明の別の一態様として、本発明の抽出物を含有することを特徴とする生物飼育用剤が提供される。
これらの発明において、生物とはたとえば養殖動物、ペット動物などであり、養殖動物としては家畜、実験動物、家禽、魚類、甲殻類または貝類が例示される。飼料としては体調の維持および／または改善用飼料が例示される。生物飼育用剤としては浸漬用剤、飼料添加剤、飲料用添加剤が例示される。
これらの発明によれば、それらを適用する前記例示するような生物において、本発明に使用される本発明の抽出物のインスリン様作用、アルドースレダクターゼ阻害作用に基づき、本発明の前記治療剤または予防剤によるのと同様の効果の発現が期待できる。すなわち、本発明の飼料等は、当該生物におけるインスリン量またはインスリン応答の変調を伴う疾患や糖尿病の合併症の治療または予防効果を発揮し得る。
本発明に使用される本発明の抽出物は通常、対象生物の体重１ｋｇ、１日当たり好ましくは０．０１〜２０００ｍｇ投与される。投与は、たとえば、当該有効成分を、対象生物に供する人工配合飼料の原料中に添加混合しておくか、人工配合飼料の粉末原料と混合した後、その他の原料にさらに添加混合することで行うことができる。また、本発明の抽出物の飼料中の含有量は特に限定されるものではなく、目的に応じて適宜設定すれば良いが、例えば前記有効成分の乾燥重量で０．００１〜１５重量％の割合が好適である。生物飼育用剤における本発明の有効成分の含有量も同程度とすればよい。
本発明の飼料の製造法に特に限定はなく、また配合も一般の飼料に準ずるものであればよく、製造された飼料中に本発明の抽出物が含まれていればよい。生物飼育用剤も同様にして調製することができる。
本発明が適用できる生物としては限定はないが、養殖動物としては、ウマ、ウシ、ブタ、ヒツジ、ヤギ、ラクダ、ラマなどの家畜、マウス、ラット、モルモット、ウサギなどの実験動物、ニワトリ、アヒル、七面鳥、駝鳥などの家禽、ペット動物としてはイヌ、ネコなどが挙げられ、広く適用できる。
本発明においては、例えば、本発明の抽出物を含んでなる飼料を摂取させること、または本発明の抽出物の含有液（例えば、前記浸漬用剤を水に溶解させたもの）に対象生物を浸漬することにより、家畜、実験動物、家禽、ペット動物などの体調を良好に維持し、または、改善させることができる。なお、これらの態様は本発明における生物の飼育方法の一態様である。
また、本発明は前記有効成分を含む細胞へのグルコース取り込み促進剤を提供することもできる。当該グルコース取り込み促進剤としては、前記有効成分そのものであってもよく、また、前記有効成分を含む組成物であってもよい。当該グルコース取り込み促進剤は、たとえば、前記有効成分を当該有効成分と同じ用途に使用可能な他の成分、たとえば、公知のインスリン製剤、インスリン分泌促進剤、インスリン抵抗性改善剤、食後過血糖改善剤、インスリン様作用剤などと配合し、上記治療剤または予防剤の製造方法に準じて通常使用される試薬の形態に製造すればよい。当該グルコース取り込み促進剤における前記有効成分の含有量は、当該グルコース取り込み促進剤の投与方法、使用目的などを考慮し、本発明の所望の効果の発現が得られ得るような量であればよく、特に限定されるものではない。典型的なグルコース取り込み促進剤中の有効成分の含有量としては、乾燥重量で０．１〜１００重量％程度である。また、該グルコース取り込み促進剤の使用量も、本発明の所望の効果の発現が得られ得る量であれば特に限定されるものではない。特に、生体に投与して使用する場合には、好ましくは前記治療剤または予防剤における有効成分の投与量の範囲内で有効成分を投与できるような量で使用すればよい。当該グルコース取り込み促進剤は、治療又は予防に細胞へのグルコース取り込み促進作用を要する疾患の治療又は予防に有用である。当該疾患としては、例えば上記のインスリン様作用を要する疾患のほか、心臓疾患、特に心筋梗塞、虚血後の心臓損傷等が例示される。また、当該グルコース取り込み促進剤は、細胞によるグルコースの取り込みを促進することから、筋肉細胞においては当該作用が機能することにより、筋肉増強作用、疲労回復作用を誘発することができる。また、当該グルコース取り込み促進剤は、これらの疾患に対する治療又は予防用の食品、飲料又は飼料の製造に使用することもできる。これらの食品、飲料又は飼料については、本発明の前述の食品、飲料又は飼料に準じて使用することができる。また、当該グルコース取り込み促進剤は上記の治療又は予防に細胞へのグルコース取り込み促進作用を要する疾患に対する薬物のスクリーニングにも有用である。さらに当該グルコース取り込み促進剤は、細胞によるグルコース取り込み作用のメカニズム研究やその細胞の物理的変化等の機能研究にも有用である。
また、本発明は前記有効成分を含む脂肪細胞への分化誘導剤を提供することもできる。当該分化誘導剤が脂肪細胞に分化誘導できる前駆細胞としては、脂肪細胞に分化しうる細胞であれば特に限定はないが、例えば前駆脂肪細胞の他、繊維芽細胞や間葉系幹細胞等が挙げられる。当該分化誘導剤としては、前記有効成分そのものであってもよく、また、前記有効成分を含む組成物であってもよい。当該分化誘導剤は、たとえば、前記有効成分を当該有効成分と同じ用途に使用可能な他の成分、たとえば、公知のインスリン製剤、インスリン分泌促進剤、インスリン抵抗性改善剤、食後過血糖改善剤、インスリン様作用剤などと配合し、上記治療剤または予防剤の製造方法に準じて通常使用される試薬の形態に製造すればよい。当該分化誘導剤における前記有効成分の含有量は、当該分化誘導剤の投与方法、使用目的などを考慮し、本発明の所望の効果の発現が得られ得るような量であればよく、特に限定されるものではない。典型的な脂肪細胞への分化誘導剤中の有効成分の含有量としては、乾燥重量で０．１〜１００重量％程度である。また、当該分化誘導剤の使用量も、本発明の所望の効果の発現が得られ得る量であれば特に限定されるものではない。特に、生体に投与して使用する場合には、好ましくは前記治療剤または予防剤における有効成分の投与量の範囲内で有効成分を投与できるような量で使用すればよい。当該分化誘導剤は、治療又は予防に脂肪細胞への分化誘導作用を要する疾患の治療又は予防に有用である。当該疾患としては、例えば上記のインスリン様作用を要する疾患のほか、痛風、脂肪肝、胆石症、月経異常、不妊症等が例示される。また、当該分化誘導剤は、これらの疾患に対する治療又は予防用の食品、飲料又は飼料の製造に使用することもできる。これらの食品、飲料又は飼料については、本発明の前述の食品、飲料又は飼料に準じて使用することができる。また、当該分化誘導剤は上記の治療又は予防に脂肪細胞への分化誘導剤を要する疾患に対する薬物のスクリーニングにも有用である。さらに当該分化誘導剤は、脂肪細胞への分化誘導作用のメカニズム研究やその物理的変化等の機能研究にも有用である。
また、本発明は前記有効成分を含むアルドースレダクターゼ阻害剤を提供することもできる。当該阻害剤としては、前記有効成分そのものであってもよく、また、前記有効成分を含む組成物であってもよい。当該阻害剤は、たとえば、前記有効成分を当該有効成分と同じ用途に使用可能な他の成分、例えばエパルレスタットなどと配合し、上記治療剤または予防剤の製造方法に準じて通常使用される試薬の形態に製造すればよい。当該阻害剤における前記有効成分の含有量は、当該阻害剤の投与方法、使用目的などを考慮し、本発明の所望の効果の発現が得られ得るような量であればよく、特に限定されるものではない。典型的なアルドースレダクターゼ阻害剤中の有効成分の含有量としては、乾燥重量で０．１〜１００重量％程度である。また、当該阻害剤の使用量も、本発明の所望の効果の発現が得られ得る量であれば特に限定されるものではない。特に、生体に投与して使用する場合には、好ましくは前記治療剤または予防剤における有効成分の投与量の範囲内で有効成分を投与できるような量で使用すればよい。当該阻害剤は、治療又は予防にアルドースレダクターゼ阻害作用を要する疾患の治療又は予防に有用である。当該疾患としては、例えば前述の糖尿病の合併症が例示される。また、当該阻害剤は、これらの疾患に対する治療又は予防用の食品、飲料又は飼料の製造に使用することもできる。これらの食品、飲料又は飼料については、前述の本発明の食品、飲料又は飼料に準じて使用することができる。また、当該阻害剤は上記の糖尿病の合併症に対する薬物のスクリーニングにも有用である。さらに当該阻害剤は、ポリオール代謝や糖尿病の合併症のメカニズム研究やその物理的変化等の機能研究にも有用である。
また、本発明の抽出物は化粧料素材としても使用することができる。該抽出物は、化粧料として使用可能な任意の物質と混合して使用可能である。一般的には、水、アルコール類、油脂類、脂肪酸類、グリセロール、無機塩類、防腐剤、界面活性剤、ビタミン類、アミノ酸類、糖類等と混合し、ローション、乳液、クリーム等として使用できる。化粧料中の本発明の抽出物の含有量としては、乾燥重量で０．１〜８０重量％程度が好適である。
また、本発明の別の態様として、キサントアンゲロールＨ、イソババカルコン、ババクロマノール、ムンドゥレアフラバノンＡ、イソババチン及びプロストラトールＦからなる群より選択される少なくとも１つ以上を有効成分として含有することを特徴とするアルドースレダクターゼ阻害剤が提供される。これらの有効成分の製造方法としては公知の方法により得ることができる。また、当該アルドースレダクターゼ阻害剤の製造方法としては、前述の本発明の治療剤又は予防剤の製造に従って製造することができる。
本発明で使用される前記有効成分は、その作用発現にとっての有効量の投与を生体に行っても毒性は認められない。たとえば経口投与の場合、アシタバの６０（ｖ／ｖ）％エタノール水溶液抽出物の乾燥物をそれぞれ１ｇ／ｋｇ体重でマウスに単回投与しても死亡例は認められない。また、前記有効成分は、ラットへの経口投与において１ｇ／ｋｇ体重を経口単回投与しても死亡例は認められない。In the present invention, a celery family plant is a plant belonging to the angiosperm family Aceraceae, and examples thereof include Ashitaba, Seri, Mitsuba, Shishiudo, carrot, celery, parsley and the like. In the present invention, Ashitaba can be particularly preferably used. In addition, the Apiaceae plant used in the present invention is not particularly limited, but fruits, seeds, seed coats, flowers, leaves, stems, roots, rhizomes and / or whole plants can be used as they are.
As a raw material for the extract of the present invention, a processed product is used in addition to the above-mentioned Apiaceae plant. Although there is no limitation in particular as a processed material here, For example, the ground material of a ciraceae plant, a shredded material, a dried material, a juice, etc. can be used. Moreover, the extract obtained by making a celery family plant into a tea leaf shape by a well-known method, and performing the extraction method of this invention on this can also be used as an extract of this invention. Moreover, as a celery family plant used as a raw material, it may be a plurality of celery family plants or a mixture of processed products thereof. In the present invention, extracts derived from different plants, extracts derived from different parts of the derived plants, extracts derived from the same, but extracted from hydroalcohols having different compositions, or Extracts obtained by combinations thereof can be used alone or in combination of two or more as the extract of the present invention. Moreover, if the extract of this invention is contained, what contains the extract obtained by the extraction method different from the manufacturing method of the extract of this invention can also be used as an extract of this invention.
The pulverized product of the family Aceraceae can be obtained, for example, by drying the plant and pulverizing it using a pulverizer, or by freeze pulverization. The shredded product can be obtained by cutting a celery family plant finely as appropriate, and the dried product can be obtained by freeze-drying, for example. The squeezed liquid can be obtained by squeezing celery family plants using a known plant squeezing method, for example, a screw type, gear type, cutter type or other squeezing machine or juicer.
In the present invention, the water-containing alcohol is not particularly limited, but refers to a mixture (alcohol aqueous solution) of alcohols such as ethanol, methanol, isopropyl alcohol, ethylene glycol, butylene glycol, propylene glycol, glycerol, and water. Particularly preferred is water-containing ethanol. The hydrous alcohol used in the present invention is not particularly limited because a larger amount of a physiologically active substance can be contained in the obtained extract. For example, the alcohol content is preferably 40 (v / v) at that concentration. % To less than 100 (v / v)%, more preferably 45 to 80 (v / v)%, particularly preferably 50 to 70 (v / v)% hydrous alcohol is used. Usually, it is appropriate to use an aqueous ethanol solution having a concentration of 40 (v / v)% or more and less than 100 (v / v)% as the hydrous alcohol. Moreover, these water-containing alcohols can be used alone or as water-containing alcohols in which various alcohols are appropriately mixed as desired.
In the present invention, an extract refers to a substance obtained through a step of performing an extraction operation using an extraction solvent. The extraction can be performed as follows by a known extraction method. For example, the extraction can be carried out in a batch or continuous manner using a solvent as it is, or after pulverizing or chopping the plant as a specific part of the ciraceae plant as a raw material. The amount of the extraction solvent may be determined as appropriate, but it is usually 0% of the weight of the raw material plant as it is in use (for example, a raw plant if the raw plant is a raw plant), preferably 0. 0.1 to 100 times the amount of extraction solvent may be used. The extraction temperature may be appropriately determined according to the purpose, but is preferably 0 to 80 ° C, more preferably 0 to 60 ° C, further preferably 0 to 50 ° C, and particularly preferably 0 to 40 ° C. It is a range. The extraction time may be determined in consideration of the extraction efficiency. Usually, the raw material, the extraction solvent, and the extraction temperature are set so as to be preferably in the range of several seconds to several days, more preferably 5 minutes to 24 hours. Is preferred. The extraction operation may be performed, for example, while stirring or standing, and may be repeated several times as necessary. By the above operation, an extract derived from a celery family plant (hereinafter sometimes referred to as the extract of the present invention) is obtained. If necessary, the extract may be further concentrated by subjecting it to treatment such as filtration, centrifugation, concentration, ultrafiltration, molecular sieving and the like.
For example, when the extract of the present invention obtained as a raw material is Ashitaba, which is a kind of Aceraceae plant, 4-hydroxyderricin or xanthoangelol, which is a chalcone compound having physiological activity, is used. It is a novel extract containing a high concentration of two chalcone compounds (hereinafter referred to as TB1 and TB2) found by the present inventors. Heretofore, there has been no report that an extract has been obtained from an Aceraceae plant using TB1 or TB2 at a high concentration as an index, and it has been carried out for the first time by the present inventors. The chemical structures of TB1 and TB2 are shown in the following formula (Formula 1) and the following formula (Formula 2), respectively.

The above four chalcone compounds (4-hydroxyderricin, xanthoangelol, TB1, TB2) contained in the extract of the present invention are naturally derived insulin-like compounds as described in Reference Examples 1 and 2 below. Active ingredient. Furthermore, it has been found by the present inventors that TB1 and TB2 also have an aldose reductase inhibitory action and are expected to have an effect on complications of diabetes (International Publication No. 2004/2004/2004). 031165 pamphlet).
Further, in the extract of the present invention obtained from Ashitaba, the chalcone compounds xanthoangelol H, isobaba chalcone, babachromanol, and flavanone compounds mundurea flavanone A, isobabatin, prostratol F These compounds have aldose reductase inhibitory action and insulin-like action as shown in Example 11 and Reference Examples 1 and 2 described later.
That is, the extract obtained by the present invention has a preventive effect on diabetes in healthy people, an improvement effect on people with early diabetes called diabetic reserve, and diabetes in people who have symptoms of complications in diabetic patients. It is extremely useful as a pharmaceutical for comprehensive improvement or prevention of diabetes and a functional food material having an effect of improving the complications. Insulin-like action of the extract of the present invention is shown in Examples 5 to 7 described later, using as an index the action of promoting glucose uptake into cells and the action of inducing differentiation into adipocytes. Moreover, the aldose reductase inhibitory action of the extract of the present invention is shown in Examples 8 and 9 described later.
In addition to the aldose reductase inhibitory action, TB1 and TB2 have a neuronal cell protective action, an NO production inhibitory action, an interleukin production inhibitory action, an osteogenesis promoting action, etc. 2004/031165 pamphlet), the extract of the present invention is also useful as a naturally-derived pharmaceutical and health food material utilizing these physiological activities.
In the present invention, the insulin-like action is not particularly limited as long as it exhibits at least one of the physiological activities of insulin, and includes, for example, promotion of sugar and amino acid uptake in cells, glycogen, protein synthesis and degradation. At least one of metabolic regulation effects such as inhibition is exemplified. The presence or absence of an insulin-like action can be easily measured by the method described in Example 5 or 6 described later. Since the active ingredient of the present invention has an insulin-like action, it can exert a therapeutic effect or a preventive effect for all diseases for which the use of insulin is effective for treatment or prevention.
The extract of the present invention also includes a fraction obtained by fractionating an extract derived from the celery family of the present invention by a known method and a fraction obtained by repeating the fractionation operation a plurality of times. The Examples of the fractionation means include extraction, fractional precipitation, column chromatography, thin layer chromatography and the like.
In the present invention, the shape of the extract of the present invention is not particularly limited, but may be any of powder, solid, and liquid. In the case of powder, there is no particular limitation, but by concentrating the extract of the present invention extracted from the raw material using hydrous alcohol as a solvent, further adding excipients and the like, drying, pulverizing, The extract of the present invention can be obtained. Moreover, the granular solid substance obtained by granulating the said extract by a well-known method can also be used as an extract of this invention. The granulation method is not particularly limited, but rolling granulation, stirring granulation, fluidized bed granulation, air flow granulation, extrusion granulation, compression molding granulation, pulverization granulation, spray granulation, spray granulation or spray granulation Examples are grains. Examples of liquid extracts include hydrous ethanol extracts themselves, concentrates and dilutions thereof, and those obtained by dissolving the powdery extract in a liquid such as water or alcohol. The
Moreover, although this invention provides the foodstuff, drink, or feed which contains the extract of this invention in high concentration or high purity, these are the foodstuff, drink, or feed of this invention compared with the conventional foodstuff, drink, or feed. It means that the chalcone compound or flavanone compound is contained in the feed in high concentration and / or high purity.
In the present invention, the extract of the present invention is referred to as the active ingredient of the present invention, and a therapeutic agent for diseases associated with modulation of insulin amount or insulin response and / or diabetic complications containing the active ingredient of the present invention or The prophylactic agent is sometimes referred to as the therapeutic agent or prophylactic agent of the present invention. In addition to the therapeutic agent and the preventive agent, the pharmaceutical agent of the present invention may be referred to as an insulin-like agent.
The active ingredient according to the present invention is not particularly toxic as will be described later. Moreover, there is no worry of side effects. Therefore, the disease can be treated or prevented safely and appropriately. Therefore, the therapeutic agent, preventive agent, food, beverage or feed of the present invention comprising the active ingredient is effective for the treatment or prevention of diseases associated with modulation of insulin amount or insulin response and / or complications of diabetes. is there.
In the present invention, diseases involving modulation of insulin amount or insulin response include changes in blood insulin level, changes in the level of insulin or insulin receptor activity, abnormalities in downstream signals of insulin receptor, and combinations thereof Diseases characterized by factors selected from, for example, diabetes, obesity, hypertension, arteriosclerosis, cocaine withdrawal symptoms, congestive heart failure, amnesia, cardiovascular spasm, cerebral vasospasm, pheochromocytoma Examples include ganglion neuroblastoma, Huntington's disease, and hyperlipidemia. Examples of diabetes include both type I diabetes and type II diabetes. In addition, type II diabetes includes diseases caused by insulin resistance, for which no therapeutic effect is observed even when insulin or an insulin secretagogue is administered.
Diseases involving modulation of insulin amount or insulin response often have insufficient insulin production or insufficient insulin action due to insulin resistance at the onset stage. Since the active ingredient of the present invention can suppress the onset of a disease accompanied by modulation of the amount of insulin or the insulin response by showing an insulin-like action, it can also be expected to have a preventive effect on the disease, and thus a complication of diabetes. A preventive effect can also be expected.
In the insulin resistance state, signals from the insulin receptor by insulin are inhibited, and various functions of insulin are not exhibited, and various metabolic abnormalities occur. The active ingredient used in the present invention can exert an insulin-like effect on symptoms of insulin resistance. That is, by using the prophylactic or therapeutic agent of the present invention, even for diseases caused by insulin resistance, for example, type II diabetes in which no therapeutic effect is observed even when insulin or an insulin secretagogue is administered. It can exert a therapeutic or preventive effect. In addition, the active ingredient of the present invention can also exert the effect of reducing the blood insulin level. That is, the medicament of the present invention can also be used as a therapeutic or prophylactic agent for diseases that require a decrease in the amount of insulin for treatment or prevention. The disease is not particularly limited, and examples thereof include hyperinsulinemia and Alzheimer's disease. In addition, since there is a report that there is a close relationship between stimulation via insulin receptor and life-prolonging effect (Science, vol. 299, P572-574 (2003); Nature, vol. 424, P277-284 (2003)). Year)), the medicament of the present invention can also be used as an anti-aging agent.
Insulin is known to promote the induction of differentiation of preadipocytes into adipocytes, and mature adipocytes are known to take up glucose and accumulate triglycerides in the cells (J. Biol Chem., Vol. 253, No. 20, P7570-7578 (1978)). That is, by utilizing this finding, the insulin-like action of the test substance can be measured by administering a test substance instead of insulin and measuring the differentiation into adipocytes and the amount of triglyceride in the cell.
Insulin is known to promote glucose uptake into cells, and in mature adipocytes, it is known that glucose uptake into cells is promoted by the action of insulin (J. Biol. Chem. , Vol. 253, No. 20, P7579-7583 (1978)). That is, using this finding, the insulin-like action of the test substance can be measured by administering a test substance instead of insulin and measuring the amount of glucose taken into mature adipocytes.
In the present invention, complications of diabetes include, for example, diabetic retinopathy, diabetic peripheral neuropathy, cataract, hearing loss, sensory abnormalities, diabetic psychosis such as muscle atrophy, diabetic nephropathy such as renal failure, arteries Examples include diabetic vascular diseases such as sclerosis and cerebral infarction, infectious diseases caused by reduced phagocytic action of leukocytes, diabetic coma, and the like.
Examples of the therapeutic agent or prophylactic agent of the present invention include those prepared by combining the active ingredient according to the present invention with a known pharmaceutical carrier. Further, as the therapeutic agent or prophylactic agent of the present invention, the active ingredient can be used for other components that can be used for the same purpose as the active ingredient, for example, known insulin preparations, insulin secretagogues, insulin resistance improving agents, postprandial It can also be blended with hyperglycemia improving agents, insulin-like agents, aldose reductase inhibitors and the like.
The therapeutic agent or prophylactic agent of the present invention is usually produced by blending the active ingredient with a pharmaceutically acceptable liquid or solid carrier, and optionally a solvent, a dispersant, an emulsifier, a buffer, Add agents, excipients, binders, disintegrants, lubricants, etc. to form solids such as tablets, granules, powders, powders, capsules, etc., and liquids such as normal solutions, suspensions, emulsions, etc. be able to. Moreover, it can also be used as a dried product which can be made liquid by adding an appropriate carrier before use, and other external preparations.
The pharmaceutical carrier can be selected according to the administration form and dosage form of the therapeutic agent or prophylactic agent. In the case of an oral preparation comprising a solid composition, it can be a tablet, pill, capsule, powder, fine granule, granule, etc., for example, starch, lactose, sucrose, mannitol, carboxymethylcellulose, corn starch Inorganic salts are used. In preparation of the oral preparation, a binder, a disintegrant, a surfactant, a lubricant, a fluidity promoter, a corrigent, a colorant, a fragrance and the like can be further added. For example, in the case of a tablet or pill, if desired, it may be coated with a sugar coating such as sucrose, gelatin or hydroxypropylcellulose, or a film of a gastric or enteric substance. In the case of an oral preparation comprising a liquid composition, it can be a pharmacologically acceptable emulsion, solution, suspension, syrup, etc. For example, purified water, ethanol, etc. are used as a carrier. Is done. Further, if desired, auxiliary agents such as wetting agents and suspending agents, sweetening agents, flavoring agents, preservatives and the like may be added.
On the other hand, in the case of a parenteral preparation, distilled water for injection, physiological saline, aqueous glucose solution, vegetable oil for injection, sesame oil, peanut oil, soybean oil, corn oil as the diluent is used as the active ingredient of the present invention according to a conventional method. It can be prepared by dissolving or suspending in propylene glycol, polyethylene glycol or the like, and adding a bactericidal agent, stabilizer, isotonic agent, soothing agent, etc., if necessary. In addition, a solid composition can be produced and dissolved in sterile water or a sterile solvent for injection before use.
External preparations include solid, semi-solid or liquid preparations for transdermal administration or transmucosal (oral or intranasal) administration. Also included are suppositories and the like. For example, emulsions such as emulsions and lotions, tinctures for external use, liquid preparations such as liquids for transmucosal administration, ointments such as oily ointments and hydrophilic ointments, transdermal administration such as films, tapes, and poultices Or a patch for transmucosal administration.
Each of the above-mentioned various preparations can be appropriately produced by a conventional method using a known pharmaceutical carrier. Further, the content of the active ingredient in such a preparation is not particularly limited as long as the active ingredient can be administered within the dosage range described below, taking into consideration its administration form, administration method and the like. . The content of the active ingredient in the therapeutic agent or prophylactic agent of the present invention is usually about 0.1 to 100% by weight.
The therapeutic agent or prophylactic agent of the present invention is administered by an appropriate administration route according to the preparation form. There is no particular limitation on the administration method, and it can be used for internal use, external use and injection. An injection can be administered, for example, intravenously, intramuscularly, subcutaneously, intracutaneously, etc. For an external preparation, for example, a suppository may be administered by its suitable administration method.
The dose as the therapeutic agent or prophylactic agent of the present invention is appropriately set according to the formulation form, administration method, purpose of use, and age, weight, and symptom of the patient to whom the therapeutic agent or prophylactic agent is administered, and is not constant. In general, the dose of the active ingredient contained in the preparation is dry weight, preferably 0.001 mg to 10 g / kg body weight, more preferably 0.1 mg to 1 g / kg body weight per day for an adult. is there. Of course, since the dosage varies depending on various conditions, an amount smaller than the above dosage may be sufficient or may be necessary beyond the range. Administration may be carried out in a single dose or divided into several doses within a desired dose range. The administration period is also arbitrary. Further, the therapeutic agent or prophylactic agent of the present invention can be administered orally as it is, or can be added to any food or drink and taken daily.
The present invention can also provide an insulin-like agent containing the active ingredient. The insulin-like agent may be the active ingredient itself or a composition containing the active ingredient. The insulin-like agent includes, for example, other components that can be used for the same use as the active ingredient, for example, known insulin preparations, insulin secretion promoters, insulin resistance improving agents, postprandial hyperglycemia improving agents. In addition, it may be mixed with an insulin-like agent and the like, and may be produced in the form of a reagent that is usually used according to the method for producing the therapeutic agent or prophylactic agent. The content of the active ingredient in the insulin-like agent may be an amount that allows the desired effect of the present invention to be obtained in consideration of the administration method, purpose of use, etc. of the insulin-like agent, It is not particularly limited. The content of the active ingredient in a typical insulin-like agent is about 0.1 to 100% by weight. Further, the amount of the insulin-like agent used is not particularly limited as long as the desired effect of the present invention can be obtained. In particular, when it is used after being administered to a living body, it is preferably used in such an amount that the active ingredient can be administered within the dose range of the active ingredient in the therapeutic agent or prophylactic agent. Insulin-like agents are useful in diseases involving modulation of insulin amount or insulin response. The insulin-like agent is also useful for screening drugs for diseases associated with modulation of insulin amount or insulin response. Further, the insulin-like agent is useful for studying the mechanism of action of insulin on cells and for functional studies relating to physical changes in the cells. In addition, the insulin-like agent can be used in addition to serum or insulin preparations, or together with them, added to a culture medium for cell / tissue / organ culture. The medium is very useful for use as a medium for cell, tissue, or organ culture that does not contain or contain serum or insulin preparations.
In addition, by administering the insulin-like agent of the present invention to humans, a decrease in blood insulin level can be expected. That is, the insulin-like agent of the present invention can also be used as a therapeutic or prophylactic agent for diseases that require a decrease in the amount of insulin for treatment or prevention. The disease is not particularly limited, and examples thereof include hyperinsulinemia and Alzheimer's disease. In addition, since there is a report that there is a close relationship between stimulation via insulin receptor and life-prolonging effect (Science, vol. 299, P572-574 (2003); Nature, vol. 424, P277-284 (2003)). Year))), the insulin-like agent of the present invention can also be used as an anti-aging agent.
The active ingredient according to the present invention is not particularly toxic as will be described later. Moreover, there is no worry of side effects. Therefore, an insulin-like action and / or an aldose reductase inhibitory action can be safely and appropriately caused in vivo. Therefore, the medicament, food, beverage or feed of the present invention comprising the active ingredient is effective for the treatment or prevention of diseases accompanied by modulation of insulin amount or insulin response and / or complications of diabetes.
Moreover, this invention provides the foodstuff, drink, or feed containing the extract of this invention. The food, beverage or feed of the present invention is extremely useful for improving or preventing symptoms of diseases associated with modulation of insulin amount or insulin response and diabetes complications due to its insulin-like action and aldose reductase inhibitory action. Furthermore, the food or beverage of the present invention is a food or beverage for lowering blood glucose levels, which has an action of lowering blood glucose levels, and is effective for those who are concerned about blood sugar levels or those who are concerned about body fat. It is useful as a functional food or beverage.
The food, beverage or feed of the present invention is another substance known to have an antidiabetic action, for example, a known insulin-like action substance, a substance having an insulin secretion promoting action, a substance having an action to improve insulin resistance Further, it can be combined with a substance having an effect of improving postprandial hyperglycemia, an aldose reductase inhibitor, and the like. For example, it can also be blended with indigestible dextrin and the like.
In the food, beverage or feed of the present invention, “containing” means containing, adding and / or diluting. Here, “containing” means that the extract of the present invention is contained in food, beverage or feed, and “adding” means adding the extract of the present invention to the raw material of food, beverage or feed. In the embodiment, “dilution” refers to an embodiment in which a food, beverage or feed ingredient is added to the extract of the present invention.
There is no particular limitation on the method for producing the food, beverage or feed of the present invention. For example, blending, cooking, processing and the like may be in accordance with those of general foods, beverages or feeds, and can be produced by the production method thereof, and the obtained foods, beverages or feeds contain the extract of the present invention. It only has to be done.
The food or beverage of the present invention is not particularly limited. For example, processed cereal products containing the extract of the present invention (processed flour products, processed starch products, premix processed products, noodles, macaronis, Breads, bean paste, buckwheat, rice cakes, rice noodles, harsame, packaging rice cakes), processed oils and fats (plastic oils, tempura oil, salad oil, mayonnaises, dressings, etc.), processed soybean products (tofu, miso, natto, etc.) ), Processed meat products (ham, bacon, press ham, sausage, etc.), marine products (frozen groundnut, kamaboko, chikuwa, hanpen, fried fish, tsumire, streaks, fish ham, sausage, bonito, processed fish eggs, canned fish , Tsukuda boiled, etc.), dairy products (raw milk, cream, yogurt, butter, cheese, condensed milk, powdered milk, ice cream, etc.), processed vegetables and fruits (pae) Tomatoes, jams, pickles, fruit drinks, vegetable drinks, mixed drinks, etc.), confectionery (chocolate, biscuits, confectionery breads, cakes, candy sweets, rice confectionery, etc.), alcoholic drinks (Japanese sake, Chinese sake, wine) , Whiskey, shochu, vodka, brandy, gin, rum, beer, soft alcoholic beverage, fruit liquor, liqueur, etc.), favorite beverage (green juice, green tea, tea, oolong tea, coffee, soft drink, lactic acid beverage, etc.), seasoning Ingredients (soy sauce, sauce, vinegar, mirin, etc.), canned / bottled / bag foods (beef rice, kettle rice, red rice, curry, other cooked foods), semi-dried or concentrated foods (lever paste, other spreads, Buckwheat noodle soup, concentrated soup), dried food (instant noodles, instant curry, instant coffee, powdered juice, powdered soup, Soba miso soup, cooked food, cooked beverage, cooked soup, etc., frozen food (sukiyaki, steamed rice bowl, eel cabbage, hamburg steak, shumai, dumplings, various sticks, fruit cocktails, etc.), solid food, liquid food (soup, etc.) ), Processed agricultural and forestry products such as spices, processed livestock products, processed fishery products and the like.
The food of the present invention contains the extract of the present invention alone or in plural, added and / or diluted, and there is no particular limitation on the shape thereof, and it is taken orally in the form of tablets, granules, capsules, etc. Possible shapes are also included.
The tablet-like food production method can be produced by a known method, and is not particularly limited. For example, Ashitaba dry powder is extracted with a 60 (v / v)% ethanol aqueous solution, and then concentrated to an extract. Additives such as dextrin, indigestible dextrin, corn starch, tapioca starch, cyclodextrin and the like, freeze-dried and pulverized to obtain a dry powder of Ashitaba extract, and then lactose, crystalline cellulose, Sucrose fatty acid ester, reduced maltose, calcium phosphate, eggshell calcium, fine silicon dioxide, CMC-Ca, trehalose, vitamin C and other vitamins, citric acid, glucose, sugar, sugar alcohol, stevia and other sweeteners, flavors, powder Fruit juice, seaweed-derived powder (eg, fucoidan-containing powder, agarooligosaccharide Powder), mushroom powder, dried vegetable powder (for example, acetaceae plant powder such as Ashitaba), etc. are mixed as appropriate, granulated as necessary, and compressed by a rotary tableting machine. it can.
Moreover, as a manufacturing method of a granular food, it can manufacture by a well-known method, Although it does not specifically limit, For example, the various mixing obtained before giving to a rotary type tableting machine in the manufacturing process of said tablet-like food The resulting powder can be kneaded by adding ethanol, granulated by an extrusion granulator, dried, and sized with a vibrating sieve to produce a granular food product.
Moreover, as a manufacturing method of a capsule-shaped foodstuff, it can manufacture by a well-known method, Although it does not specifically limit, For example, various mixing obtained before giving to a rotary type tableting machine in the manufacturing process of said tablet-shaped foodstuff The prepared powder can be filled into No. 1 capsules. In addition, glycerin fatty acid ester, beeswax and the like can be added to the powder before filling and emulsified to produce soft capsules using gelatin and glycerin as the encapsulating material.
In addition, the extract of the present invention has reduced bitterness and astringency unique to celery family plants, and is also excellent as a food material. That is, the food or beverage of the present invention is a food or beverage excellent in taste.
In addition, the beverage of the present invention can be produced by a known method, and is not particularly limited. For example, dried Ashitaba powder is extracted with a 60 (v / v)% ethanol aqueous solution and concentrated to an extract. Add dry form powder such as dextrin, indigestible dextrin, corn starch, tapioca starch, cyclodextrin, etc., freeze-dry and grind to obtain dry powder of Ashitaba extract. Add ingredients, fruit juice, powdered fruit juice, vegetable extract, vegetable paste, seaweed-derived powder or seaweed extract (eg, fucoidan-containing material, agaro-oligosaccharide-containing material, etc.), mushroom powder, dried powder of celery family, etc. The drink of this invention can be manufactured by melt | dissolving in alcohol. Moreover, after melt | dissolving the dry powder of an Ashitaba extract in water, the drink of this invention can also be manufactured according to the manufacturing method of a well-known soft drink. Moreover, the concentrate of an Ashitaba extract can also be used instead of the dry powder of an Ashitaba extract. Furthermore, an alcoholic beverage can also be manufactured as a drink of this invention using the water-containing alcoholic extract of Ashitaba.
The content of the active ingredient in the food or beverage of the present invention is not particularly limited and can be appropriately selected from the viewpoint of its functionality and activity expression. For example, the dry weight of the active ingredient in the food, preferably 0. 00001% by weight or more, more preferably 0.0001 to 90% by weight, and even more preferably 0.0006 to 80% by weight. For example, in beverages, preferably 0.00001% by weight or more, more preferably 0.0001. It is -90 weight%, More preferably, it is 0.0006-80 weight%. The food or beverage of the present invention is ingested so that the active ingredient contained therein is, for example, preferably 0.001 mg to 10 g / kg body weight, more preferably 0.1 mg to 1 g / kg body weight per day for an adult. do it.
The present invention also provides a biological feed comprising, adding and / or diluting the extract of the present invention. Furthermore, as another aspect, the extract of the present invention is added to a living organism. There is also provided a method of raising an organism characterized by administration. Moreover, as another aspect of the present invention, there is provided a biological breeding agent comprising the extract of the present invention.
In these inventions, organisms are, for example, farm animals, pet animals, etc., and examples of farm animals include domestic animals, laboratory animals, poultry, fish, shellfish or shellfish. Examples of the feed include a feed for maintaining and / or improving physical condition. Examples of biological breeding agents include soaking agents, feed additives, and beverage additives.
According to these inventions, the therapeutic agent or prophylaxis of the present invention based on the insulin-like action and aldose reductase inhibitory action of the extract of the present invention used in the present invention in the organisms exemplified above to which they are applied. The same effect as that produced by the agent can be expected. That is, the feed or the like of the present invention can exert an effect of treating or preventing a disease accompanied by modulation of insulin amount or insulin response in the organism or a complication of diabetes.
The extract of the present invention used in the present invention is usually administered in an amount of 1 kg body weight of the target organism, preferably 0.01 to 2000 mg per day. The administration is performed, for example, by adding and mixing the active ingredient into the raw material of the artificial blended feed to be provided to the target organism, or by mixing with the powder raw material of the artificial blended feed and further adding and mixing to the other raw materials. be able to. Further, the content of the extract of the present invention in the feed is not particularly limited, and may be appropriately set according to the purpose. For example, the ratio of 0.001 to 15% by weight based on the dry weight of the active ingredient Is preferred. The content of the active ingredient of the present invention in the biological breeding agent may be set to the same level.
There is no particular limitation on the method for producing the feed of the present invention, and the composition may be the same as that of general feed, and the extract of the present invention may be contained in the produced feed. The biological breeding agent can be prepared in the same manner.
The organisms to which the present invention can be applied are not limited, but farmed animals include horses, cattle, pigs, sheep, goats, camels, llamas and other domestic animals, mice, rats, guinea pigs, rabbits and other laboratory animals, chickens, ducks. Examples of poultry such as turkeys and eagle birds, and pet animals include dogs and cats, which can be widely applied.
In the present invention, for example, a feed comprising the extract of the present invention is ingested, or the target organism is placed in a liquid containing the extract of the present invention (for example, the above-mentioned soaking agent dissolved in water). By soaking, the physical condition of livestock, laboratory animals, poultry, pet animals, etc. can be maintained well or improved. In addition, these aspects are one aspect | mode of the biological breeding method in this invention.
Moreover, this invention can also provide the glucose uptake | capture promoter to the cell containing the said active ingredient. The glucose uptake promoter may be the active ingredient itself or a composition containing the active ingredient. The glucose uptake promoter is, for example, other components that can be used for the same use as the active ingredient, such as a known insulin preparation, an insulin secretion promoter, an insulin resistance improving agent, a postprandial hyperglycemia improving agent. In addition, it may be mixed with an insulin-like agent and the like, and may be produced in the form of a reagent that is usually used according to the method for producing the therapeutic agent or prophylactic agent. The content of the active ingredient in the glucose uptake promoter may be an amount that allows the desired effect of the present invention to be obtained in consideration of the administration method, purpose of use, etc. of the glucose uptake promoter, It is not particularly limited. The content of the active ingredient in a typical glucose uptake promoter is about 0.1 to 100% by weight in dry weight. Further, the amount of the glucose uptake promoter used is not particularly limited as long as the desired effect of the present invention can be obtained. In particular, when it is used after being administered to a living body, it is preferably used in such an amount that the active ingredient can be administered within the dose range of the active ingredient in the therapeutic agent or prophylactic agent. The glucose uptake promoter is useful for the treatment or prevention of diseases that require an action of promoting glucose uptake into cells for treatment or prevention. Examples of such diseases include heart diseases such as myocardial infarction and heart damage after ischemia in addition to the above-mentioned diseases requiring insulin-like action. Moreover, since the said glucose uptake | capture promoter accelerates | stimulates the uptake | capture of glucose by a cell, when the said function functions in a muscle cell, a muscle reinforcement | strengthening effect | action and a fatigue recovery effect | action can be induced. Moreover, the said glucose uptake | capture promoter can also be used for manufacture of the foodstuff, drink, or feed for the treatment or prevention with respect to these diseases. About these foodstuffs, drinks, or feed, it can use according to the above-mentioned foodstuff, drink, or feed of this invention. Moreover, the said glucose uptake | capture promoter is useful also for the screening of the drug with respect to the disease which requires the glucose uptake | capture promotion effect | action to a cell for said treatment or prevention. Further, the glucose uptake promoter is useful for studying the mechanism of glucose uptake by cells and for functional studies such as physical changes in the cells.
Moreover, this invention can also provide the differentiation-inducing agent to the adipocyte containing the said active ingredient. The precursor cells that can be differentiated into adipocytes by the differentiation inducer are not particularly limited as long as they are cells that can differentiate into adipocytes. For example, in addition to precursor adipocytes, fibroblasts, mesenchymal stem cells and the like can be mentioned. It is done. The differentiation inducer may be the active ingredient itself or a composition containing the active ingredient. The differentiation-inducing agent is, for example, other components that can be used for the same use as the active ingredient, for example, known insulin preparations, insulin secretion promoters, insulin resistance improving agents, postprandial hyperglycemia improving agents, What is necessary is just to mix | blend with an insulin-like agent etc. and to manufacture in the form of the reagent normally used according to the manufacturing method of the said therapeutic agent or preventive agent. The content of the active ingredient in the differentiation-inducing agent is not particularly limited as long as the desired effect of the present invention can be obtained in consideration of the administration method, purpose of use, etc. of the differentiation-inducing agent. Is not to be done. The content of the active ingredient in the agent for inducing differentiation into typical adipocytes is about 0.1 to 100% by weight in dry weight. Further, the amount of the differentiation inducer used is not particularly limited as long as the desired effect of the present invention can be obtained. In particular, when it is used after being administered to a living body, it is preferably used in such an amount that the active ingredient can be administered within the dose range of the active ingredient in the therapeutic agent or prophylactic agent. The differentiation inducer is useful for the treatment or prevention of diseases that require an action of inducing differentiation into adipocytes for treatment or prevention. Examples of the disease include gout, fatty liver, cholelithiasis, menstrual abnormalities, infertility, and the like in addition to the above-mentioned diseases requiring insulin-like action. Moreover, the said differentiation inducer can also be used for manufacture of the food, drink, or feed for the treatment or prevention with respect to these diseases. About these foodstuffs, drinks, or feed, it can use according to the above-mentioned foodstuff, drink, or feed of this invention. The differentiation inducer is also useful for screening drugs for diseases that require an adipocyte differentiation inducer for the treatment or prevention described above. Furthermore, the differentiation inducer is useful for studying the mechanism of the differentiation-inducing action on adipocytes and functional studies such as physical changes.
Moreover, this invention can also provide the aldose reductase inhibitor containing the said active ingredient. The inhibitor may be the active ingredient itself or a composition containing the active ingredient. The inhibitor is, for example, a combination of the active ingredient and other ingredients that can be used for the same purpose as the active ingredient, for example, epalrestat, etc. What is necessary is just to manufacture to a form. The content of the active ingredient in the inhibitor is not particularly limited as long as the desired effect of the present invention can be obtained in consideration of the administration method and purpose of use of the inhibitor. It is not a thing. The content of the active ingredient in a typical aldose reductase inhibitor is about 0.1 to 100% by weight in dry weight. Further, the amount of the inhibitor used is not particularly limited as long as the desired effect of the present invention can be obtained. In particular, when it is used after being administered to a living body, it is preferably used in such an amount that the active ingredient can be administered within the dose range of the active ingredient in the therapeutic agent or prophylactic agent. The inhibitor is useful for treating or preventing a disease requiring an aldose reductase inhibitory action for treatment or prevention. Examples of the disease include the aforementioned complications of diabetes. Moreover, the said inhibitor can also be used for manufacture of the food, drink, or feed for the treatment or prevention with respect to these diseases. These foods, beverages or feeds can be used according to the foods, beverages or feeds of the present invention described above. The inhibitor is also useful for screening drugs for the above-mentioned diabetic complications. Furthermore, the inhibitor is also useful for functional studies such as polyol metabolism and diabetic complication mechanisms and physical changes.
The extract of the present invention can also be used as a cosmetic material. The extract can be used by mixing with any substance that can be used as a cosmetic. Generally, it can be mixed with water, alcohols, fats and oils, fatty acids, glycerol, inorganic salts, preservatives, surfactants, vitamins, amino acids, saccharides, etc., and used as a lotion, emulsion, cream or the like. As content of the extract of this invention in cosmetics, about 0.1 to 80 weight% is suitable by dry weight.
Moreover, as another aspect of the present invention, at least one selected from the group consisting of xanthoangelol H, isobaba chalcone, babachromanol, mundrea flavanone A, isobabatin and prostratol F is used as an active ingredient. An aldose reductase inhibitor characterized by containing is provided. These active ingredients can be produced by known methods. Moreover, as a manufacturing method of the said aldose reductase inhibitor, it can manufacture according to manufacture of the above-mentioned therapeutic agent or preventive agent of this invention.
The said active ingredient used by this invention does not recognize toxicity, even if it administers to a living body the effective amount for the effect | action expression. For example, in the case of oral administration, no deaths are observed even when a single dose of 1 g / kg body weight of a dried product of acitaba 60 (v / v)% aqueous ethanol solution is administered to mice. Moreover, even if the said active ingredient is orally administered to a rat and orally administers 1 g / kg body weight once orally, a death example is not recognized.

  EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated further more concretely, this invention is not limited to these description at all. Note that% in the examples means capacity (v / v)% unless otherwise specified.

40 mL of extraction solvent (40, 50, 60, 70, 80% ethanol aqueous solution and 100% ethanol) was added to 2 g of pulverized lyophilized product of Ashitaba leaf stem, and extraction was performed at 25 ° C. for 30 minutes. The supernatant after the centrifugation was quantified for chalcones using HPLC. The column used was TSK gel ODS-80Ts QA (4.6 mm × 25 cm: manufactured by Tosoh Corporation). Solvent A (mixed distilled water and acetonitrile in a volume ratio of 4: 1, containing 0.1% trifluoroacetic acid) and solvent B (mixed distilled water and acetonitrile in a volume ratio of 1: 9, 0.1 The solvent B ratio was linearly maintained at 0 to 100% until 0 to 45 minutes, and the solvent B ratio was maintained at 100% for 10 minutes. The elution rate was 1 mL / min, the detection was 215 nm, and the column temperature was 30 ° C.
Table 1 shows the results. As shown in Table 1, it has been clarified that aqueous ethanol extraction is more efficient than 100% ethanol for the extraction of TB1 and TB2 from Ashitaba leaf stem. It was also found that 60%, 70%, and 80% ethanol aqueous solutions were more efficient than 100% ethanol for extraction of xanthoangelol and 4-hydroxyderricin from Ashitaba leaf stems.

実施例３ アシタバ葉茎部抽出物の調製
アシタバ葉茎部の凍結乾燥物を粉砕したもの２ｇに、４０ｍＬの抽出溶媒（４０、５０、６０、７０、８０％エタノール水溶液ならびに１００％エタノール）を加え、室温で３０分間抽出を行った。遠心後の上清３０ｍＬを濃縮乾固し０．７５ｍＬのジメチルスルホキシドに溶解した。
実施例４ アシタバ根部抽出物の調製
アシタバ根部の凍結乾燥物を粉砕したもの２ｇに、４０ｍＬの抽出溶媒（４０、５０、６０、７０、８０％エタノール水溶液ならびに１００％エタノール）を加え、室温で３０分間抽出を行った。遠心後の上清３０ｍＬを濃縮乾固し０．７５ｍＬのジメチルスルホキシドに溶解した。
実施例５ アシタバ葉茎部抽出物の脂肪細胞への分化誘導活性
（１）脂肪細胞への分化誘導
脂肪細胞への分化誘導はＲｕｂｉｎ Ｃ．Ｓ．らの方法（Ｊ．Ｂｉｏｌ．Ｃｈｅｍ．，Ｖｏｌ．２５３，Ｎｏ．２０，ｐ７５７０〜７５７８（１９７８年））を一部改良して行った。２００μＭアスコルビン酸を含む１０％ウシ胎児血清（ギブコ社製）含有ダルベッコ改良イーグル培地（シグマ社製，Ｄ６０４６）（以下Ａ−Ｄ−ＭＥＭ培地）に前駆脂肪細胞株３Ｔ３−Ｌ１（ＡＴＣＣ ＣＣＬ−９２．１）を４×１０^３個／ｍＬになるように懸濁し、１２穴マイクロタイタープレートのウェルに２ｍＬずつ加えて５％炭酸ガス存在下、３７℃で７日間培養した。なお、２，４日目に同培地により培地交換を行った。７日目に、０．２５μＭデキサメタゾンを含むＡ−Ｄ−ＭＥＭ培地に交換後、各ウエルそれぞれ実施例３で調製したアシタバ葉茎部４０％エタノール水溶液抽出物ジメチルスルホキシド溶液を終濃度０．１％、０．０３３％となるように、５０％エタノール水溶液抽出物ジメチルスルホキシド溶液を終濃度０．１％、０．０３３％、０．０１１％となるように、また６０％、７０％、８０％エタノール水溶液抽出物あるいは１００％エタノール抽出物ジメチルスルホキシド溶液を終濃度０．２％、０．０６７％、０．０２２％となるように添加した。なお陽性対照として４μＬの５ｍｇ／ｍＬインスリン（タカラバイオ社製）水溶液添加の区分を、陰性対照としてジメチルスルホキシド添加の区分を設定した。４５時間後にＡ−Ｄ−ＭＥＭ培地に交換し、各ウエルそれぞれアシタバ葉茎部４０％エタノール水溶液抽出物ジメチルスルホキシド溶液を終濃度０．１％、０．０３３％となるように、５０％エタノール水溶液抽出物ジメチルスルホキシド溶液を終濃度０．１％、０．０３３％、０．０１１％となるように、また６０％、７０％、８０％エタノール水溶液抽出物あるいは１００％エタノール抽出物ジメチルスルホキシド溶液を終濃度０．２％、０．０６７％、０．０２２％となるように添加し、陽性対照として２μＬの５ｍｇ／ｍＬインスリン水溶液、陰性対照としてジメチルスルホキシドを添加し，さらに７日間培養した。なお、２、４日目に培地を交換し、その際各ウェルに、アシタバ葉茎部４０％エタノール水溶液抽出物ジメチルスルホキシド溶液を終濃度０．１％、０．０３３％となるように、５０％エタノール水溶液抽出物ジメチルスルホキシド溶液を終濃度０．１％、０．０３３％、０．０１１％となるように、また６０％、７０％、８０％エタノール水溶液抽出物あるいは１００％エタノール抽出物ジメチルスルホキシド溶液を終濃度０．２％、０．０６７％、０．０２２％となるように添加した。陽性対照として２μＬの５ｍｇ／ｍＬインスリン水溶液、陰性対照としてジメチルスルホキシドを添加した。
（２）トリグリセリド生合成量の測定
成熟脂肪細胞への分化誘導の指標として、またインスリン様作用の評価として細胞中のトリグリセリドの量を測定した。培養終了後、培地を除き、リン酸緩衝塩溶液で２回細胞を洗浄し、１ｍＬのヘキサン：イソプロパノール＝３：２の溶媒を添加して３０分間室温に置いた後上清を回収した。この操作を再度繰り返し、得られた２ｍＬの上清を濃縮乾固した。沈殿を１００μＬのイソプロパノールに溶解後、溶液１０μＬ中に含まれるトリグリセリドの量をトリグリセライドＥ−テスト（和光純薬社製、ｃｏｄｅ ４３２−４０２０１）を用い測定した。また、測定は全て２連で行った。
この結果、ジメチルスルホキシド添加区分と比較してアシタバ葉茎部４０％、５０％、６０％、７０％、８０％エタノール水溶液抽出物あるいは１００％エタノール抽出物添加区分において、インスリンを添加した区分と同様にトリグリセリド生合成の誘導が確認できた。すなわち、アシタバ葉茎部４０％、５０％、６０％、７０％、８０％エタノール水溶液抽出物あるいは１００％エタノール抽出物に脂肪細胞への分化誘導活性が認められた。
実施例６ アシタバ根部抽出物によるグルコース取り込み促進作用
（１）成熟脂肪細胞の調製
成熟脂肪細胞への分化誘導は前述のＲｕｂｉｎ Ｃ．Ｓ．らの方法を一部改良し、行った。２００μＭアスコルビン酸を含む１０％ウシ胎児血清含有ダルベッコ改良イーグル培地に３Ｔ３−Ｌ１細胞を４×１０^３個／ｍＬになるように懸濁し、１２穴マイクロタイタープレートのウエルに２ｍＬずつ加えて５％炭酸ガス存在下、３７℃で７日間培養した。７日目に、２００μＭアスコルビン酸、０．２５μＭデキサメタゾン及び１０μｇ／ｍＬインスリン、０．５ｍＭ ３−イソブチル−１−メチルキサンチン（ナカライテスク社製、１９６２４−４４）を含む１０％ウシ胎児血清含有ダルベッコ改良イーグル培地２ｍＬに交換した。４５時間後に２００μＭアスコルビン酸及び５μｇ／ｍＬインスリンを含む１０％ウシ胎児血清含有ダルベッコ改良イーグル培地２ｍＬに交換し、さらに２日後、４日後に同培地を交換し７日間培養することで成熟脂肪細胞を調製した。
（２）成熟脂肪細胞へのグルコース取り込み促進作用の測定
グルコース取り込み促進作用の評価として、またインスリン様作用の評価として成熟脂肪細胞においてサンプル（実施例４で調製したアシタバ根部抽出物）刺激時の細胞内への２−デオキシグルコース取り込み量を測定した。
培養終了後、培地を除き、０．１（ｗ／ｖ）％牛血清アルブミン（シグマ社製、Ａ８０２２）含有ダルベッコ改良イーグル培地で２回細胞を洗浄した後、各ウエルそれぞれ終濃度０．１％、０．０６７％、０．０２２％の実施例４で調製したアシタバ根部４０％、５０％エタノール水溶液抽出物ジメチルスルホキシド溶液、終濃度０．２％、０．０６７％、０．０２２％のアシタバ根部６０％エタノール水溶液抽出物ジメチルスルホキシド溶液、終濃度０．０６７％、０．０２２％のアシタバ根部７０％、８０％エタノール水溶液抽出物ジメチルスルホキシド溶液あるいはアシタバ根部１００％エタノール抽出物ジメチルスルホキシド溶液を含む同培地１ｍＬを添加し、５％炭酸ガス存在下、３７℃で一晩培養した。なお、陰性対照としてサンプルを添加しない区分を設定した。一晩培養後、ヘペス緩衝塩溶液（１４０ｍＭ ＮａＣｌ、５ｍＭ ＫＣｌ、２．５ｍＭ ＭｇＳＯ_４、１ｍＭ ＣａＣｌ_２、２０ｍＭ ＨＥＰＥＳ−Ｎａ （ｐＨ ７．４））で２回細胞を洗浄し、各ウエルそれぞれ終濃度０．１％、０．０６７％、０．０２２％のアシタバ根部４０％、５０％エタノール水溶液抽出物ジメチルスルホキシド溶液、終濃度０．２％、０．０６７％、０．０２２％のアシタバ根部６０％エタノール水溶液抽出物ジメチルスルホキシド溶液、終濃度０．０６７％、０．０２２％のアシタバ根部７０％、８０％エタノール水溶液抽出物ジメチルスルホキシド溶液あるいはアシタバ根部１００％エタノール抽出物ジメチルスルホキシド溶液を含む同緩衝液０．９ｍＬを添加し、３７℃で７５分培養した。この際陽性対照として、４５分経過した時点でサンプルを添加していないウエルで終濃度１μｇ／ｍＬとなるようにインスリンを添加した区分を設定した。その後、０．５μＣｉ／ｍＬ ２−デオキシ−［１，２−^３Ｈ（Ｎ）］−グルコース（パーキンエルマーライフサイエンス社製、ＮＥＴ５４９Ａ）、１ｍＭ ２−デオキシグルコース（ナカライテスク社製、１０７２２−１１）含有ヘペス塩緩衝液１００μＬを添加しさらに３７℃で１０分培養した。培養終了後、上清を除去し、４℃に冷却したリン酸塩緩衝液で３回細胞を洗浄後、１％ノニデットＰ−４０含有リン酸塩緩衝液０．５ｍＬを添加し細胞を溶解することで、細胞中に取り込まれた２−デオキシ−［１，２−^３Ｈ（Ｎ）］−グルコースを溶出した。上清２５μＬを用いてウルチマゴールド（パーキンエルマーライフサイエンス社製、６０１３３２９）をシンチレーションカクテルとして液体シンチレーションカウンターＬＳ６５００（ベックマン社製）により放射活性を測定した。
この結果、アシタバ根部４０％、５０％、６０％、７０％、８０％エタノール水溶液抽出物あるいは１００％エタノール抽出物を添加した区分で陰性対照と比較してインスリンを添加した区分と同様に２−デオキシ［１，２−^３Ｈ（Ｎ）］−グルコース取り込みの促進が見られた。すなわち、アシタバ根部４０％、５０％、６０％、７０％、８０％エタノール水溶液抽出物あるいは１００％エタノール抽出物にグルコース取り込み促進活性が認められた。
実施例７ アシタバ根部抽出物の脂肪細胞への分化誘導
実施例４によって調製したアシタバ根部４０％、５０％、６０％、７０％、８０％エタノール水溶液抽出物あるいは１００％エタノール抽出物の成熟脂肪細胞への分化誘導作用（インスリン様作用）を実施例５の方法に準じて測定した。
すなわち、サンプルとして、各ウエルそれぞれ終濃度０．１％、０．０３３％のアシタバ根部４０％エタノール水溶液抽出物ジメチルスルホキシド溶液、終濃度０．０３３％、０．０１１％のアシタバ根部５０％エタノール水溶液抽出物ジメチルスルホキシド溶液、終濃度０．０２２％のアシタバ根部７０％エタノール水溶液抽出物ジメチルスルホキシド溶液、終濃度０．０６７％、０．０２２％のアシタバ根部６０％、８０％エタノール水溶液抽出物ジメチルスルホキシド溶液あるいは１００％エタノール抽出物ジメチルスルホキシド溶液を添加した区分を設定した。なお陽性対照として４μＬの５ｍｇ／ｍＬインスリン（タカラバイオ社製）水溶液添加の区分を、陰性対照としてジメチルスルホキシド添加の区分を設定した。この後実施例５記載の方法と同様に、培地およびサンプルの交換を行い、サンプル添加７日後に細胞中のトリグリセリドの量を測定した。
この結果、アシタバ根部４０％、５０％、６０％、７０％、８０％エタノール水溶液抽出物あるいは１００％エタノール抽出物添加区分においてトリグリセリド生合成の誘導が認められた。すなわち、アシタバ根部４０％、５０％、６０％、７０％、８０％エタノール水溶液抽出物あるいは１００％エタノール抽出物に成熟脂肪細胞への分化誘導作用が認められた。
調製例１ アシタバ葉茎部、根部水抽出物の調製
アシタバ葉茎部、および根部の凍結乾燥物を粉砕したもの２ｇに、４０ｍＬの水を加え、室温で３０分間抽出を行った。遠心後の上清３０ｍＬを濃縮乾固し０．７５ｍＬの水に溶解し、アシタバ葉茎部、根部水抽出物とした。
実施例８ アシタバ根部抽出物のアルドースレダクターゼ阻害作用
アシタバ根部抽出物のアルドースレダクターゼ阻害作用を以下の方法により測定した。サンプルとして実施例４で調製したアシタバ根部各種エタノール抽出物、および調製例１で調製したアシタバ根部水抽出物（５０％ジメチルスルホキシド水溶液に溶解したもの）１０μＬ、０．２Ｍリン酸緩衝液（ｐＨ６．２）１００μＬ、１ｍＭ ＮＡＤＰＨ（リン酸緩衝液）２０μＬ、人筋肉細胞由来アルドースレダクターゼ溶液（０．１Ｕ／ｍＬ、和光純薬工業社製、リン酸緩衝液）１０μＬに１００ｍＭメチルグリオキサール溶液２０μＬを加え、３０秒経過の後より１８０秒間、ＮＡＤＰＨの３４０ｎｍにおける吸光度の変化を測定した。陰性対照としてサンプルの代わりに５０％ジメチルスルホキシド水溶液を使用した。また、各サンプルのブランクとしてメチルグリオキサール溶液の代わりに蒸留水を使用して吸光度を測定した。測定値は２回の実験値の平均値で示した。アルドースレダクターゼ阻害率（％）は以下の式により算出した。
阻害率（％）＝［１−（△Ａｓ−△Ａｓｂ）／（△Ａｃ−△Ａｃｂ）］×１００
ここで、△Ａｓ及び△Ａｃはそれぞれサンプル溶液、陰性対照溶液の１分間あたりの吸光度変化を示し、△Ａｓｂ及び△Ａｃｂはそれぞれサンプル溶液、陰性対照溶液のブランク溶液の１分間あたりの吸光度変化を示す。
サンプルの添加量は最終濃度を表３に示す通りとした。表３に示すように、アシタバ根部含水エタノール抽出物は水抽出物やエタノール抽出物よりも高いアルドースレダクターゼ阻害作用を示すことが明らかとなった。

実施例９ アシタバ葉茎部抽出物のアルドースレダクターゼ阻害作用
実施例３で調製したアシタバ葉茎部抽出物のアルドースレダクターゼ阻害活性を実施例８と同様の方法により測定した。表４にその結果を示す。表４に示すように、アシタバ葉茎部６０％エタノール水溶液抽出物は水抽出物やエタノール抽出物よりも高いアルドースレダクターゼ阻害活性を示すことが明らかとなった。

実施例１０ キサントアンゲロールＨ、イソババカルコン、ババクロマノール、ムンドゥレアフラバノンＡ、イソババチン、プロストラトールＦの調製
実施例３で調製したアシタバ葉茎部４０、５０、６０、７０、および８０％エタノール抽出物と実施例４で調製したアシタバ根部４０、５０、６０、７０、および８０％エタノール抽出物について、実施例１に示す条件でＨＰＬＣによる分析を行った結果、これらの抽出物中にキサントアンゲロールＨ、イソババカルコン、ババクロマノール、ムンドゥレアフラバノンＡ、イソババチン、プロストラトールＦが含まれていることを確認した。これらの化合物を各種クロマトグラフィーを用いて単離した。
実施例１１ アシタバ根部抽出物に含まれる物質のアルドースレダクターゼ阻害活性
実施例１０で調製したキサントアンゲロールＨ、イソババカルコン、ババクロマノール、ムンドゥレアフラバノンＡ、イソババチン、プロストラトールＦのアルドースレダクターゼ阻害活性を実施例８と同様の方法で測定した。
表５にその結果を示す。表５に示すように、キサントアンゲロールＨ、イソババカルコン、ババクロマノール、ムンドゥレアフラバノンＡ、イソババチン、プロストラトールＦには濃度依存的にアルドースレダクターゼ阻害活性があることが明らかとなった。
実施例１および２において含水エタノール抽出物中にＴＢ１、ＴＢ２が含有されていることを確認しているが、本発明者らは国際公開第２００４／０３１１６５号パンフレットにおいてＴＢ１およびＴＢ２にアルドースレダクターゼ阻害作用があることを明らかにしている。すなわち、実施例１および２に示す含水エタノール抽出物中には、ＴＢ１、ＴＢ２以外にもアルドースレダクターゼ阻害作用を有する物質が多く含まれていることが明らかとなった。

参考例１ キサントアンゲロール、４−ハイドロキシデリシン、ＴＢ１、ＴＢ２、キサントアンゲロールＨ、イソババカルコン、ムンドゥレアフラバノンＡ、イソババチン、プロストラトールＦの脂肪細胞への分化誘導活性
キサントアンゲロール、４−ハイドロキシデリシン、ＴＢ１、ＴＢ２、キサントアンゲロールＨ、イソババカルコン、ムンドゥレアフラバノンＡ、イソババチン、プロストラトールＦの成熟脂肪細胞への分化誘導作用（インスリン様作用）を実施例５の方法に準じて測定した。
すなわち、サンプルとして、各ウエルにそれぞれキサントアンゲロール（終濃度１、３、１０μＭ）、４−ハイドロキシデリシン（終濃度３、１０μＭ）、ＴＢ１（終濃度３、１０μＭ）、ＴＢ２（終濃度３、１０μＭ）、イソババカルコン（終濃度３、１０μＭ）、キサントアンゲロールＨ（終濃度１．３、４、１３μＭ）、ムンドゥレアフラバノンＡ（終濃度３、１０μＭ）、イソババチン（終濃度３、１０、３０μＭ）、プロストラトールＦ（終濃度３、１０μＭ）のジメチルスルホキシド溶液を添加した区分を設定した。なお陽性対照として４μＬの５ｍｇ／ｍＬインスリン（タカラバイオ社製）水溶液添加の区分を、陰性対照としてジメチルスルホキシド添加の区分を設定した。この後実施例５記載の方法と同様に、培地およびサンプルの交換を行い、サンプル添加７日後に細胞中のトリグリセリドの量を測定した。
この結果、キサントアンゲロール、４−ハイドロキシデリシン、ＴＢ１、ＴＢ２、キサントアンゲロールＨ、イソババカルコン、ムンドゥレアフラバノンＡ、イソババチン、プロストラトールＦ添加区分において、それぞれトリグリセリド生合成の誘導が認められた。すなわち、キサントアンゲロール、４−ハイドロキシデリシン、ＴＢ１、ＴＢ２、キサントアンゲロールＨ、イソババカルコン、ムンドゥレアフラバノンＡ、イソババチン、プロストラトールＦに成熟脂肪細胞への分化誘導作用が認められた。
参考例２ キサントアンゲロール、４−ハイドロキシデリシン、キサントアンゲロールＨ、イソババカルコンによるグルコース取り込み促進作用
キサントアンゲロール、４−ハイドロキシデリシン、キサントアンゲロールＨ、イソババカルコンのグルコース取り込み促進作用の評価として、またインスリン様作用の評価として、実施例６記載の方法に準じて成熟脂肪細胞でのサンプル刺激時の細胞内への２−デオキシグルコース取り込み量を測定した。
すなわち、サンプルとして各ウエルそれぞれキサントアンゲロール（終濃度３、１０μＭ）、４−ハイドロキシデリシン（終濃度３、１０、３０μＭ）、キサントアンゲロールＨ（終濃度３０μＭ）、イソババカルコン（終濃度３、１０、３０μＭ）のジメチルスルホキシド溶液を用いた。なお、陰性対照としてサンプルを添加しない区分を、陽性対照として、終濃度１μｇ／ｍＬとなるようにインスリンを添加した区分を設定した。この後同様に、細胞中に取り込まれた２−デオキシ−［１，２−^３Ｈ（Ｎ）］−グルコースを測定した。
この結果、キサントアンゲロール、４−ハイドロキシデリシン、キサントアンゲロールＨ、イソババカルコンを添加した区分で陰性対照と比較してインスリンを添加した区分と同様に２−デオキシ［１，２−^３Ｈ（Ｎ）］−グルコース取り込みの促進が見られた。すなわち、キサントアンゲロール、４−ハイドロキシデリシン、キサントアンゲロールＨ、イソババカルコンにグルコース取り込み促進活性が認められた。The pulverized lyophilized product of Ashitaba root was extracted in the same manner as in Example 1, and the amount of chalcones in the extract was quantified. Table 2 shows the results. As shown in Table 2, it has been clarified that aqueous ethanol extraction is more efficient than 100% ethanol for extracting TB1 and TB2 from the root of Ashitaba. It was also found that 60%, 70%, and 80% ethanol aqueous solutions were more efficient than 100% ethanol for extraction of xanthoangelol and 4-hydroxyderricin from the root of Ashitaba.

Example 3 Preparation of Ashitaba leaf stem extract To 2 g of crushed lyophilized product of Ashitaba leaf stem, 40 mL extraction solvent (40, 50, 60, 70, 80% ethanol aqueous solution and 100% ethanol) was added. Extraction was performed at room temperature for 30 minutes. 30 mL of the supernatant after centrifugation was concentrated to dryness and dissolved in 0.75 mL of dimethyl sulfoxide.
Example 4 Preparation of Ashitaba Root Extract 40 g of extraction solvent (40, 50, 60, 70, 80% ethanol aqueous solution and 100% ethanol) was added to 2 g of the crushed lyophilized product of Ashitaba root, and 30 at room temperature. Extraction was performed for minutes. 30 mL of the supernatant after centrifugation was concentrated to dryness and dissolved in 0.75 mL of dimethyl sulfoxide.
Example 5 Differentiation Inducing Activity of Ashitaba Leaf Stem Extract into Adipocytes (1) Induction of Differentiation into Adipocytes Differentiation induction into adipocytes was performed by Rubin C. et al. S. The method (J. Biol. Chem., Vol. 253, No. 20, p7570-7578 (1978)) was partially improved. Pre-adipocyte cell line 3T3-L1 (ATCC CCL-92.) Was added to Dulbecco's modified Eagle medium (Sigma, D6046) (hereinafter referred to as AD-MEM medium) containing 10% fetal bovine serum (Gibco) containing 200 μM ascorbic acid. 1) was suspended at 4 × 10 ³ cells / mL, 2 mL was added to each well of a 12-well microtiter plate, and the cells were cultured at 37 ° C. for 7 days in the presence of 5% carbon dioxide gas. On the second and fourth days, the medium was replaced with the same medium. On the 7th day, the medium was replaced with AD-MEM medium containing 0.25 μM dexamethasone, and then each well was treated with a 40% ethanol aqueous solution extract of Ashitaba leaf stem prepared in Example 3 in each well. 50% ethanol aqueous solution dimethyl sulfoxide solution to a final concentration of 0.1%, 0.033%, 0.011%, and 60%, 70%, 80% Aqueous ethanol extract or 100% ethanol extract dimethyl sulfoxide solution was added to final concentrations of 0.2%, 0.067%, and 0.022%. As a positive control, 4 μL of 5 mg / mL insulin (manufactured by Takara Bio Inc.) aqueous solution was added, and as a negative control, dimethyl sulfoxide was added. After 45 hours, the medium was replaced with AD-MEM medium, and each well had a 50% ethanol aqueous solution so that the final concentration of 0.1% and 0.033% dimethyl sulfoxide solution was obtained from each of Ashitaba leaf stem 40% ethanol aqueous solution. Extract dimethyl sulfoxide solution to a final concentration of 0.1%, 0.033%, 0.011%, 60%, 70%, 80% aqueous ethanol extract or 100% ethanol extract dimethyl sulfoxide solution. The final concentrations were 0.2%, 0.067% and 0.022%, 2 μL of 5 mg / mL insulin aqueous solution was added as a positive control, and dimethyl sulfoxide was added as a negative control, followed by further culturing for 7 days. On the second and fourth days, the medium was changed, and each well was treated with 50% ethanol aqueous solution extract dimethyl sulfoxide solution of Ashitaba leaves and stems at a final concentration of 0.1% and 0.033%. % Ethanol aqueous extract dimethyl sulfoxide solution to a final concentration of 0.1%, 0.033%, 0.011%, 60%, 70%, 80% ethanol aqueous extract or 100% ethanol extract dimethyl The sulfoxide solution was added to final concentrations of 0.2%, 0.067%, and 0.022%. 2 μL of a 5 mg / mL insulin aqueous solution was added as a positive control, and dimethyl sulfoxide was added as a negative control.
(2) Measurement of triglyceride biosynthesis The amount of triglyceride in cells was measured as an index for inducing differentiation into mature adipocytes and as an evaluation of insulin-like action. After completion of the culture, the medium was removed, the cells were washed twice with a phosphate buffered saline solution, 1 mL of hexane: isopropanol = 3: 2 solvent was added, and the mixture was allowed to stand at room temperature for 30 minutes, and then the supernatant was collected. This operation was repeated again, and the resulting 2 mL supernatant was concentrated to dryness. After dissolving the precipitate in 100 μL of isopropanol, the amount of triglyceride contained in 10 μL of the solution was measured using a triglyceride E-test (manufactured by Wako Pure Chemical Industries, code 432-40201). All measurements were performed in duplicate.
As a result, compared to the dimethyl sulfoxide addition category, the Ashitaba leaf stem portion 40%, 50%, 60%, 70%, 80% ethanol aqueous solution extract or 100% ethanol extract addition category is the same as the category to which insulin is added. Induction of triglyceride biosynthesis was confirmed. That is, differentiation induction activity into adipocytes was observed in 40%, 50%, 60%, 70%, 80% ethanol aqueous extract or 100% ethanol extract of Ashitaba leaf stem.
Example 6 Glucose Uptake Promoting Action by Ashitaba Root Extract (1) Preparation of Mature Adipocytes Differentiation into mature adipocytes was induced by the aforementioned Rubin C.I. S. Some of these methods were improved. Suspend 3T3-L1 cells in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum containing 200 μM ascorbic acid to 4 × 10 ³ cells / mL, add 2 mL to each well of a 12-well microtiter plate, and add 5% carbonic acid. The cells were cultured at 37 ° C. for 7 days in the presence of gas. On day 7, 10% fetal bovine serum-containing Dulbecco's improvement containing 200 μM ascorbic acid, 0.25 μM dexamethasone and 10 μg / mL insulin, 0.5 mM 3-isobutyl-1-methylxanthine (Nacalai Tesque, 19624-44) The eagle medium was replaced with 2 mL. After 45 hours, replace with 2 mL of Dulbecco's modified Eagle's medium containing 10% fetal bovine serum containing 200 μM ascorbic acid and 5 μg / mL insulin, and after 2 days and 4 days, the medium is replaced and cultured for 7 days. Prepared.
(2) Measurement of glucose uptake promoting action on mature adipocytes Cells upon stimulation of sample (Ashitaba root extract prepared in Example 4) in mature adipocytes as an evaluation of glucose uptake promotion action and as an insulin-like action The amount of 2-deoxyglucose uptake was measured.
After completion of the culture, the medium was removed, and the cells were washed twice with Dulbecco's modified Eagle's medium containing 0.1 (w / v)% bovine serum albumin (Sigma, A8022), and each well had a final concentration of 0.1%. 0.067%, 0.022% Ashitaba root 40% prepared in Example 4, 50% aqueous ethanol extract dimethyl sulfoxide solution, final concentrations 0.2%, 0.067%, 0.022% Ashitaba 60% ethanol aqueous solution extract dimethyl sulfoxide solution, final concentration 0.067%, 0.022% Ashitaba root 70%, 80% ethanol aqueous solution dimethyl sulfoxide solution or Ashitaba root 100% ethanol extract dimethyl sulfoxide solution 1 mL of the same medium was added and cultured overnight at 37 ° C. in the presence of 5% carbon dioxide gas. In addition, the category which does not add a sample was set as a negative control. After overnight culture, the cells were washed twice with Hepes buffered salt solution (140 mM NaCl, 5 mM KCl, 2.5 mM MgSO ₄ , 1 mM CaCl ₂ , 20 mM HEPES-Na (pH 7.4)), and each well had a final concentration. 0.1%, 0.067%, 0.022% Ashitaba root 40%, 50% aqueous ethanol extract dimethyl sulfoxide solution, final concentration 0.2%, 0.067%, 0.022% Ashitaba root 60 The same buffer containing 70% ethanol solution extract dimethyl sulfoxide, final concentration 0.067%, 0.022% ashitaba root 70%, 80% ethanol aqueous solution dimethyl sulfoxide solution or ashitaba root 100% ethanol extract dimethyl sulfoxide solution 0.9 mL of the solution was added and cultured at 37 ° C. for 75 minutes. At this time, as a positive control, a section in which insulin was added was set so that the final concentration was 1 μg / mL in a well in which no sample was added after 45 minutes. Thereafter, 0.5 μCi / mL 2-deoxy- [1,2- ³ H (N)]-glucose (manufactured by PerkinElmer Life Sciences, NET549A), 1 mM 2-deoxyglucose (manufactured by Nacalai Tesque, 10722-11) 100 μL of the Hepes salt buffer solution was added, and further cultured at 37 ° C. for 10 minutes. After completion of the culture, the supernatant is removed, the cells are washed three times with a phosphate buffer cooled to 4 ° C., and 0.5 mL of 1% nonidet P-40-containing phosphate buffer is added to lyse the cells. As a result, 2-deoxy- [1,2- ³ H (N)]-glucose incorporated into the cells was eluted. Using 25 μL of the supernatant, radioactivity was measured with a liquid scintillation counter LS6500 (Beckman) using Ultima Gold (Perkin Elmer Life Sciences, 6013329) as a scintillation cocktail.
As a result, in the group to which 40%, 50%, 60%, 70%, 80% ethanol aqueous extract or 100% ethanol extract was added as the root of Ashitaba, compared to the negative control, 2- Promotion of deoxy [1,2- ³ H (N)]-glucose uptake was observed. That is, glucose uptake promoting activity was observed in 40%, 50%, 60%, 70%, 80% aqueous ethanol extract or 100% ethanol extract of Ashitaba root.
Example 7 Induction of differentiation of Ashitaba root extract into adipocytes Mature adipocytes of Acitaba root 40%, 50%, 60%, 70%, 80% ethanol aqueous solution extract or 100% ethanol extract prepared according to Example 4 Differentiation inducing action (insulin-like action) was measured according to the method of Example 5.
That is, as a sample, each well has a final concentration of 0.1%, 0.033% Ashitaba root 40% ethanol aqueous solution extract, a dimethyl sulfoxide solution, a final concentration of 0.033%, 0.011% Ashitaba root 50% ethanol aqueous solution. Extract dimethyl sulfoxide solution, final concentration 0.022% Ashitaba root 70% ethanol aqueous solution extract dimethyl sulfoxide solution, final concentration 0.067%, 0.022% Ashitaba root 60%, 80% aqueous ethanol extract dimethyl sulfoxide A section to which a solution or a 100% ethanol extract dimethyl sulfoxide solution was added was set. As a positive control, 4 μL of 5 mg / mL insulin (manufactured by Takara Bio Inc.) aqueous solution was added, and as a negative control, dimethyl sulfoxide was added. Thereafter, in the same manner as described in Example 5, the medium and the sample were exchanged, and the amount of triglyceride in the cells was measured 7 days after the addition of the sample.
As a result, induction of triglyceride biosynthesis was observed in the sections where 40%, 50%, 60%, 70%, 80% aqueous ethanol extract or 100% ethanol extract was added. That is, 40%, 50%, 60%, 70%, 80% aqueous ethanol extract or 100% ethanol extract of Ashitaba root was found to induce differentiation into mature adipocytes.
Preparation Example 1 Preparation of Ashitaba leaf stem and root water extract 40 mL of water was added to 2 g of the crushed freeze dried product of Ashitaba leaf stem and root, and extraction was performed at room temperature for 30 minutes. 30 mL of the supernatant after centrifugation was concentrated and dried and dissolved in 0.75 mL of water to obtain an extract of Ashitaba leaf stem and root water.
Example 8 Aldose Reductase Inhibitory Action of Ashitaba Root Extract The aldose reductase inhibitory action of Ashitaba root extract was measured by the following method. Samples of various ethanol extracts of Ashitaba root prepared in Example 4 and Acitaba root water extract prepared in Preparation Example 1 (dissolved in 50% aqueous dimethyl sulfoxide solution) 10 μL, 0.2 M phosphate buffer (pH 6. 2) 100 μL, 1 mM NADPH (phosphate buffer) 20 μL, human muscle cell-derived aldose reductase solution (0.1 U / mL, Wako Pure Chemical Industries, Ltd., phosphate buffer) 10 μL, 100 mM methylglyoxal solution 20 μL is added, The change in absorbance at 340 nm of NADPH was measured for 180 seconds after 30 seconds had elapsed. A 50% aqueous dimethyl sulfoxide solution was used instead of the sample as a negative control. Moreover, the absorbance was measured using distilled water instead of the methylglyoxal solution as a blank for each sample. The measured value was shown as the average of two experimental values. The aldose reductase inhibition rate (%) was calculated by the following formula.
Inhibition rate (%) = [1- (ΔAs−ΔAsb) / (ΔAc−ΔAcb)] × 100
Here, ΔAs and ΔAc indicate the change in absorbance per minute for the sample solution and the negative control solution, respectively, and ΔAsb and ΔAcb indicate the change in absorbance per minute for the blank solution of the sample solution and the negative control solution, respectively. Show.
The amount of sample added was as shown in Table 3 for the final concentration. As shown in Table 3, it was revealed that Ashitaba root hydrous ethanol extract showed higher aldose reductase inhibitory action than water extract and ethanol extract.

Example 9 Aldose reductase inhibitory action of Ashitaba leaf stem extract The aldose reductase inhibitory activity of the Ashitaba leaf stem extract prepared in Example 3 was measured by the same method as in Example 8. Table 4 shows the results. As shown in Table 4, it was clarified that the 60% ethanol aqueous extract of Ashitaba leaf stem part showed higher aldose reductase inhibitory activity than the water extract and ethanol extract.

Example 10 Preparation of Xanthoangelol H, Isobaba Chalcone, Babachromanol, Mundrea Flavanone A, Isobabachin, Prostratol F Ashitaba Leaf Stem 40, 50, 60, 70, and 80 Prepared in Example 3 % Ethanol extract and Ashitaba root 40, 50, 60, 70, and 80% ethanol extract prepared in Example 4 were analyzed by HPLC under the conditions shown in Example 1. As a result, It was confirmed that xanthoangelol H, isobaba chalcone, babachromanol, Mundrea flavanone A, isobabatin and prostratol F were contained. These compounds were isolated using various chromatographies.
Example 11 Aldose Reductase Inhibitory Activity of Substances Included in Ashitaba Root Extract Xanthoangelol H, Isobaba chalcone, Babachromanol, Mundrea flavanone A, Isobabatin, Prostratol F aldose reductase prepared in Example 10 Inhibitory activity was measured in the same manner as in Example 8.
Table 5 shows the results. As shown in Table 5, it was revealed that xanthoangelol H, isobaba chalcone, babachromanol, Mundurea flavanone A, isobabatin and prostratol F have aldose reductase inhibitory activity in a concentration-dependent manner. .
In Examples 1 and 2, it was confirmed that TB1 and TB2 were contained in the water-containing ethanol extract. However, in the WO 2004/031165 pamphlet, the present inventors inhibited aldose reductase on TB1 and TB2. It is clear that there is. That is, it has been clarified that the water-containing ethanol extract shown in Examples 1 and 2 contains many substances having an aldose reductase inhibitory action in addition to TB1 and TB2.

Reference Example 1 Xanthoangelol, 4-hydroxyderricin, TB1, TB2, xanthoangelol H, isobaba chalcone, Mundrea flavanone A, isobabatin, prostratol F differentiation induction activity into adipocytes Xanthoangel Example of action of inducing differentiation (insulin-like action) of roll, 4-hydroxyderricin, TB1, TB2, xanthoangelol H, isobaba chalcone, mundrea flavanone A, isobabatin and prostratol F into mature adipocytes Measurement was carried out according to the method of No. 5.
That is, as a sample, xanthoangelol (final concentration 1, 3, 10 μM), 4-hydroxyderricin (final concentration 3, 10 μM), TB1 (final concentration 3, 10 μM), TB2 (final concentration 3) were added to each well. 10 μM), isobaba chalcone (final concentration 3, 10 μM), xanthoangelol H (final concentration 1.3, 4, 13 μM), Mundrea flavanone A (final concentration 3, 10 μM), isobabatin (final concentration 3) 10, 30 μM), and a dimethyl sulfoxide solution of prostratol F (final concentration 3, 10 μM) was added. As a positive control, 4 μL of 5 mg / mL insulin (manufactured by Takara Bio Inc.) aqueous solution was added, and as a negative control, dimethyl sulfoxide was added. Thereafter, in the same manner as described in Example 5, the medium and the sample were exchanged, and the amount of triglyceride in the cells was measured 7 days after the addition of the sample.
As a result, xanthoangelol, 4-hydroxyderricin, TB1, TB2, xanthoangelol H, isobaba chalcone, mundrea flavanone A, isobabatin, and prostratol F were added, respectively. Admitted. That is, xanthoangelol, 4-hydroxyderricin, TB1, TB2, xanthoangelol H, isobaba chalcone, mundrea flavanone A, isobabatin, and prostratol F are recognized to induce differentiation into mature adipocytes. It was.
Reference Example 2 Glucose uptake promoting action by xanthoangelol, 4-hydroxyderricin, xanthoangelol H, isobaba chalcone Xanthoangelol, 4-hydroxyderricin, xanthoangelol H, glucose of isobaba chalcone As an evaluation of the uptake promoting action and an evaluation of the insulin-like action, the amount of 2-deoxyglucose uptake into the cells upon stimulation of the sample with mature adipocytes was measured according to the method described in Example 6.
That is, xanthoangelol (final concentration 3, 10 μM), 4-hydroxyderricin (final concentration 3, 10, 30 μM), xanthoangelol H (final concentration 30 μM), isobaba chalcone (final concentration) as a sample. A dimethyl sulfoxide solution having a concentration of 3, 10, 30 μM) was used. In addition, the division which added the sample as a negative control and the division which added insulin so that it might become a final concentration of 1 microgram / mL were set as a positive control. Thereafter, similarly, 2-deoxy- [1,2- ³ H (N)]-glucose incorporated into the cells was measured.
As a result, in the group to which xanthoangelol, 4-hydroxyderricin, xanthoangelol H, isobabachalcone was added, compared to the negative control, 2-deoxy [1,2- ³ H (N)]-glucose uptake was promoted. That is, glucose uptake promoting activity was observed in xanthoangelol, 4-hydroxyderricin, xanthoangelol H, and isobaba chalcone.

  40 kg of dried Ashitaba powder was added to 400 L of 55% ethanol and extracted at 25 ° C. for 1 hour. Thereafter, the Ashitaba powder was separated by filtration, and the extract was concentrated under reduced pressure using a concentrating can to obtain a 150 L Ashitaba extract concentrate. To this concentrate, 10 kg of dextrin was added as an excipient, freeze-dried and pulverized to obtain 22 kg of Ashitaba extract powder.

  Into the mixer, 2.5 kg of Ashitaba extract powder obtained in Example 12, 10.0 kg of crystalline cellulose, and 0.6 kg of sucrose fatty acid ester were sequentially added and stirred for 15 minutes. This mixture was tableted with a rotary tableting machine to obtain 13.1 kg of tablets. In addition, tablets of Ashitaba dry powder were produced in the same manner except that the Ashitaba dry powder described in Example 12 was used as a control. When these sensory tests were conducted by 20 panel members, it was evaluated that the tablets containing the extract of Ashitaba had better bitterness and astringency and were superior in taste.

  5.0 kg of Ashitaba extract powder obtained in Example 12, 5.0 kg of crystalline cellulose, 5.0 kg of lactose, and 0.7 kg of sucrose fatty acid ester were sequentially added to the mixer and stirred for 15 minutes. This mixture was tableted using a rotary tableting machine to obtain 15.7 kg of tablets. In addition, tablets of Ashitaba dry powder were produced in the same manner except that the Ashitaba dry powder described in Example 12 was used as a control. When these sensory tests were conducted by 20 panel members, it was evaluated that the tablets containing the Ashitaba extract had reduced bitterness and astringency and were superior in taste.

  Into the mixer, 2.5 kg of Ashitaba extract powder obtained in Example 12, 10.0 kg of crystalline cellulose, and 0.6 kg of sucrose fatty acid ester were sequentially added and stirred for 15 minutes. Furthermore, 60% ethanol aqueous solution was added and kneaded, and the resulting mixture was granulated by an extrusion granulator. Further, this was dried at 60 ° C. for 6 hours with a shelf-type hot air dryer, and sized with a vibrating sieve to obtain 13.1 kg of a 20-100 mesh granule. In addition, granules of Ashitaba dry powder were produced in the same manner except that the Ashitaba dry powder described in Example 12 was used as a control. When these sensory tests were conducted by 20 panel members, it was evaluated that the granular product containing the extract of Ashitaba had a lower bitterness and astringency and was superior in taste.

  In a mixer, 2.0 kg of Ashitaba extract powder obtained in Example 12, 3.0 kg of lactose, 5.0 kg of trehalose, 3.5 kg of crystalline cellulose, 0.8 kg of sucrose fatty acid ester, 0.5 kg of vitamin C, citric acid 0. After sequentially charging 2 kg, the mixture was stirred for 15 minutes. Furthermore, 60% ethanol aqueous solution was added and kneaded, and granulated by an extrusion granulator. Further, this was dried at 60 ° C. for 6 hours with a shelf-type hot air dryer, and sized with a vibrating sieve to obtain 15 kg of granules of 20 to 100 mesh. In addition, granules of Ashitaba dry powder were produced in the same manner except that the Ashitaba dry powder described in Example 12 was used as a control. When these sensory tests were conducted by 20 panel members, it was evaluated that the granular product containing the extract of Ashitaba had a lower bitterness and astringency and was superior in taste.

  2.9 kg of Ashitaba extract powder obtained in Example 12, 10.78 kg of lactose, 0.9 kg of sucrose fatty acid ester, 0.1 kg of vitamin C, 0.07 kg of talc, and 0.25 kg of fragrance were sequentially added to the mixer and stirred for 15 minutes. 15 kg of mixed powder was obtained. Thereafter, 300 mg of this mixed powder was filled into No. 1 capsules to obtain capsules containing Ashitaba extract powder.

  To the blender, 10.0 kg of soybean oil was added as a base oil, and 2.0 kg of Ashitaba extract powder obtained in Example 12, 1.0 kg of vitamin E, 1.0 kg of glycerin fatty acid ester, and 0.8 kg of beeswax were added and emulsified. The obtained emulsion was filled into soft capsules containing gelatin and glycerin as encapsulating materials.

表６の材料を順次混合し、均一化させた後、プレートヒーターで９５℃、１５秒間加熱殺菌し、５０ｍＬ容量のガラス瓶に充填した。その後、パストライザーにより更に７５℃、１５分間の殺菌を行った。
また、対照として実施例１２に記載のアシタバ乾燥粉末を使用する以外は同様の方法でアシタバ乾燥粉末含有清涼飲料水を製造した。パネルメンバー２０名によりこれらの官能検査を行ったところ、アシタバ抽出物含有清涼飲料水の方が苦味も渋味も低減されており、嗜好的に優れているとの評価であった。Ashitaba extract-containing soft drink was prepared. The formulation is shown in Table 6.

The materials in Table 6 were sequentially mixed and homogenized, and then heat sterilized with a plate heater at 95 ° C. for 15 seconds, and filled into a 50 mL glass bottle. Thereafter, the pasteurizer was further sterilized at 75 ° C. for 15 minutes.
Moreover, the dry drink containing an Ashitaba powder was manufactured by the same method except using the dried Ashitaba powder described in Example 12 as a control. When these sensory tests were performed by 20 panel members, the bitterness and astringency were reduced in the Ashitaba extract-containing soft drink, and it was evaluated that the taste was superior.

表７の材料を順次混合し、均一化させた後、プレートヒーターで９５℃、１５秒間加熱殺菌し、５０ｍＬ容量のガラス瓶に充填した。その後、パストライザーにより更に７５℃、１５分間の殺菌を行った。
また、対照として実施例１２に記載のアシタバ乾燥粉末を使用する以外は同様の方法でアシタバ乾燥粉末含有清涼飲料水を製造した。パネルメンバー２０名によりこれらの官能検査を行ったところ、アシタバ抽出物含有清涼飲料水の方が苦味も渋味も低減されており、嗜好的に優れているとの評価であった。Ashitaba extract-containing soft drink was prepared. The formulation is shown in Table 7.

The materials in Table 7 were sequentially mixed and homogenized, and then heat sterilized with a plate heater at 95 ° C. for 15 seconds, and filled into a 50 mL glass bottle. Thereafter, the pasteurizer was further sterilized at 75 ° C. for 15 minutes.
Moreover, the dry drink containing an Ashitaba powder was manufactured by the same method except using the dried Ashitaba powder described in Example 12 as a control. When these sensory tests were performed by 20 panel members, the bitterness and astringency were reduced in the Ashitaba extract-containing soft drink, and it was evaluated that the taste was superior.

表８の材料を順次混合し、均一化させた後、プレートヒーターで９８℃、１５秒間加熱殺菌し、２００ｍＬ容量の缶に１９０ｇ充填した後、パストライザーにより更に８５℃、５分間の殺菌を行った。
また、対照として実施例１２に記載のアシタバ乾燥粉末を使用する以外は同様の方法でアシタバ乾燥粉末含有清涼飲料水を製造した。パネルメンバー２０名によりこれらの官能検査を行ったところ、アシタバ抽出物含有清涼飲料水の方が苦味も渋味も低減されており、嗜好的に優れているとの評価であった。Ashitaba extract-containing soft drink was prepared. The formulation is shown in Table 8.

The materials in Table 8 were mixed in order and homogenized, and then heat-sterilized with a plate heater at 98 ° C for 15 seconds. After filling 190 g into a 200 mL capacity can, further pasteurized at 85 ° C for 5 minutes. It was.
Moreover, the dry drink containing an Ashitaba powder was manufactured by the same method except using the dried Ashitaba powder described in Example 12 as a control. When these sensory tests were performed by 20 panel members, the bitterness and astringency were reduced in the Ashitaba extract-containing soft drink, and it was evaluated that the taste was superior.

表９の材料を順次混合し、均一化させた後、プレートヒーターで９５℃、１５秒間加熱殺菌し、５０ｍＬ容量のガラス瓶に充填した。その後、パストライザーにより更に７５℃、１５分間の殺菌を行った。
また、対照として実施例１２に記載のアシタバ乾燥粉末を使用する以外は同様の方法でアシタバ乾燥粉末含有清涼飲料水を製造した。パネルメンバー２０名によりこれらの官能検査を行ったところ、アシタバ抽出物含有清涼飲料水の方が苦味も渋味も低減されており、嗜好的に優れているとの評価であった。A soft drink using the Ashitaba extract concentrate obtained in Example 12 was prepared. The formulation is shown in Table 9.

The materials in Table 9 were sequentially mixed and homogenized, and then heat sterilized with a plate heater at 95 ° C. for 15 seconds, and filled into a 50 mL glass bottle. Thereafter, the pasteurizer was further sterilized at 75 ° C. for 15 minutes.
Moreover, the dry drink containing an Ashitaba powder was manufactured by the same method except using the dried Ashitaba powder described in Example 12 as a control. When these sensory tests were performed by 20 panel members, the bitterness and astringency were reduced in the Ashitaba extract-containing soft drink, and it was evaluated that the taste was superior.

  The soft drinks produced in Examples 19 to 22 were filled and sterilized in the same manner in a 200 mL capacity pouch with a stopper or a 100 mL capacity retort pouch instead of a glass bottle or a can to obtain a pouch drink.

  INDUSTRIAL APPLICABILITY According to the present invention, a hydroalcoholic extract derived from a celery family plant and a production method thereof are provided. In the extract, there are many physiologically active substances derived from natural products (for example, the substance showing the insulin-like action and aldose reductase inhibitory action, etc.), and the extract contains insulin such as diabetes or obesity. It is useful as a material for pharmaceuticals, foods, beverages or feeds that is effective for diseases involving modulation of the amount or insulin response, and / or complications of diabetes. In addition, when the food or beverage is ingested as a daily food or drink, it is possible to prevent or ameliorate symptoms associated with a disease associated with modulation of insulin amount or insulin response and / or diabetic complications. Therefore, the functional food and drink containing the extract of the present invention is a functional food and drink useful for maintaining homeostasis due to its insulin-like action and / or aldose reductase inhibitory action. In addition, the present invention also provides an insulin-like agent containing the extract of the present invention, and the insulin-like agent is useful for studying insulin functions and screening for drugs for diseases related to insulin. The present invention also provides a glucose uptake promoter for cells containing the extract of the present invention, and the glucose uptake promoter is used for the treatment or prevention of diseases that require the action of promoting glucose uptake into cells for treatment or prevention. It is also useful for the production of foods, beverages or feeds for the treatment or prevention of the disease, and for screening for drugs for diseases that require the action of promoting glucose uptake. The present invention also provides an agent for inducing differentiation into an adipocyte containing the extract of the present invention, and the agent for inducing differentiation is treatment or prevention of a disease that requires an action for inducing differentiation into an adipocyte for treatment or prevention. It is also useful for the production of foods, beverages or feeds for the treatment or prevention of the disease, and for screening for drugs for diseases that require the differentiation-inducing action. The present invention also provides an aldose reductase inhibitor containing the extract of the present invention, and the inhibitor is used for the treatment or prevention of a disease requiring an aldose reductase inhibitory action for the treatment or prevention, or the treatment or prevention of the disease. It is also useful for the production of foods, beverages or feeds, and for screening for drugs for diseases that require such inhibitory action.

Claims (13)

An extract obtained by extracting a celery family plant or a processed product thereof with hydrous alcohol. The extract according to claim 1, wherein the Apiaceae plant is Ashitaba. The extract according to claim 1 or 2, wherein the hydrous alcohol is an aqueous ethanol solution having a concentration of 40 (v / v)% or more and less than 100 (v / v)%. A method for producing an extract derived from a celery family plant or a processed product thereof, the method comprising a step of extracting the cetaceae plant or a processed product thereof with a hydrous alcohol. The method according to claim 4, wherein the Apiaceae plant is Ashitaba. The method according to claim 4 or 5, wherein the hydrous alcohol is an aqueous ethanol solution having a concentration of 40 (v / v)% or more and less than 100 (v / v)%. A therapeutic or prophylactic agent for diseases and / or diabetic complications involving modulation of insulin amount or insulin response, comprising the extract according to any one of claims 1 to 3 as an active ingredient. An insulin-like agent comprising the extract according to any one of claims 1 to 3 as an active ingredient. A food, beverage or feed comprising the extract according to any one of claims 1 to 3. The food, beverage or feed according to claim 9, which is used for treatment or prevention of diseases associated with modulation of insulin amount or insulin response and / or complications of diabetes. A glucose uptake promoter for cells containing the extract according to any one of claims 1 to 3 as an active ingredient. An agent for inducing differentiation into adipocytes, comprising the extract according to any one of claims 1 to 3 as an active ingredient. An aldose reductase inhibitor comprising the extract according to any one of claims 1 to 3 as an active ingredient.